Background technology
Erythrocyte mainly is made up of cell membrane and hemoglobin, is the important component of blood, and major function is delivering oxygen and carbon dioxide.In the higher pulmonary of partial pressure of oxygen, hemoglobin and oxygen are combined into HbO2 Oxyhemoglobin, flow to the lower tissue of partial pressure of oxygen with blood then; Hemoglobin is separated, and the oxygen that carries is discharged, and combines with the carbon dioxide that tissue is produced again; Be transported to pulmonary, excrete.Human body needs supplemental blood behind massive blood loss.Popularization along with component blood transfusion technology needn't replenish whole blood after losing blood, gets final product but give blood constitutent such as a certain amount of erythrocyte.
Therefore, erythrocytic external preservation and bioactive maintenance become the technological precursor of component blood transfusion technology.Mainly contain fluid preservation method, freeze preservation and freeze-drying with erythrocytic store method at present.Wherein liquid processes generally refers to 4 ℃ of cryopreservation methods, and freezing preservation refers to-80 ℃ or-196 ℃ of profound hypothermia preservations.Freeze-drying be with its suitably dehydration than liquid processes and freezing method remarkable advantages is arranged after drying is processed, its goods can room temperature preservation, is easy to aquation, weight alleviates greatly, is convenient to transport, and cuts the waste.At present, the fluid preservation erythrocyte has been widely used in clinical, and obtains constantly perfect; Freezing preservation erythrocyte has begun to be applied to clinical; Erythrocytic technology immaturity is still preserved in lyophilization, lacks practicality.
The research that erythrocyte is preserved is extremely paid attention to.The length of its storage life has become an important indicator weighing a national clinical blood transfusion level.Research is at present thought, adopts freeze drying technology can prolong erythrocytic storage life, and crucial adjuvants such as cryoprotective agent commonly used in the research comprise trehalose, dimethyl sulfoxide, polyvinylpyrrolidone and bovine serum albumin etc.
Trehalose is a kind of stable irreducibility disaccharidase, and its hydroxyl can replace hydrone partly to combine with protein surface, can form protective layer behind the entering cell, and pair cell forms protection.But have only when trehalose reaches finite concentration and be evenly distributed on inside and outside the cell membrane, could in freeze-drying process, play a protective role erythrocyte.Therefore, the payload trehalose is that one of chief factors of success are preserved in the erythrocyte lyophilizing to born of the same parents before the lyophilizing.Therefore yet erythrocyte membrane has impermeability to trehalose, the trehalose load is got in the erythrocyte born of the same parents, needs complicated technical process, consuming timely reaches 4 hours, and needs to introduce more kinds of chemical substances.Make technological problems and safety issue become one of difficult problem of puzzlement erythrocyte lyophilized powder application.
Dimethyl sulfoxide can prevent to form in the freeze-drying process ice crystal pair cell and cause damage.Temperature is during near 0 ℃, and dimethyl sulfoxide descends the freezing point of solution, no ice crystal formation; When temperature continues to reduce, form ice crystal in the extracellular fluid, and solution does not freeze in the erythrocyte.The outer water of erythrocyte born of the same parents constantly forms ice crystal, and born of the same parents' extracellular concentration increases, and intracellular Free water permeate through cell membranes is exuded to outside the born of the same parents, if cooling rate is enough slow, erythrocyte is dewatered gradually and can not forms ice crystal.But; Dimethyl sulfoxide is as a kind of chemical reagent; Human body is had certain toxic action, be inappropriate for drug administration by injection, residual dimethyl sulfoxide may accumulate gradually in the long-term erythrocytic patient's body of infusion lyophilizing and form injury; So need flushing repeatedly before using, thereby problem that causes the erythrocyte response rate to reduce.
Polyvinylpyrrolidone is similar with trehalose, is the impermeability cytoprotective, and FD is had the certain protection effect.Bovine serum albumin then is a kind of film protective agent.
Results of laboratory is thought at present, and in the erythrocyte freeze-drying process, needing trehalose, polyvinylpyrrolidone, bovine serum albumin combination to use could be to the effect of the performance of the erythrocyte in freeze-drying process certain protection.And needing dimethyl sulfoxide effectively to get into cell, trehalose plays a role.And, the at present various erythrocyte freeze-dry process and the prescription of research and development, because of complicated component, there are problems such as the erythrocyte response rate is low, the variation of lyophilizing red cell morphology, the active reduction of lyophilizing red blood cell enzyme in reason such as technological parameter is wayward.Therefore, seek new safely and can keep lyophilizing erythrocyte high-recovery and highly active erythrocyte lyophilized powder prescription and technology, become the key factor that has become the clinical use of restriction erythrocyte.
Tetrahydropyrimidine, chemistry by name 1,4,5; 6-tetrahydrochysene-2-methyl-4-pyrimidine carboxylic or 2-Methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid or (S)-2-methyl-1; 4,5,6-tetra-hydro pyrimidine-4-carboxylic acid or 1,4; 5,6-tetrahydro-2-methyl-4-pyrimidine carbonic acid, CAS number is 96702-03-3; Be the amino acid derivativges of finding in the marine organism in 1985, have preserve moisture, effect such as anti-ultraviolet irradiation, be used to cosmetics in recent years as moisture retention composition or adjuvant.
The specific embodiment
Below in conjunction with embodiment the present invention is done further explanation.Should be understood that following examples only are used to explain the present invention, rather than restriction protection scope of the present invention.
Embodiment 1 injection erythrocyte lyophilized powder prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Tetrahydropyrimidine 12mg
Polyvinylpyrrolidone 10mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 1 * 10 with erythrocyte
9The suspension of cell/ml adds tetrahydropyrimidine and polyvinylpyrrolidone then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 12Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 2 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Hydroxy tetrahydro pyrimidine 20mg
Polyvinylpyrrolidone 7mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds hydroxy tetrahydro pyrimidine and polyvinylpyrrolidone then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 9Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 3 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Hydroxy tetrahydro pyrimidine 30mg
Trehalose 45mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds hydroxy tetrahydro pyrimidine and trehalose then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 10Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 4 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Hydroxy tetrahydro pyrimidine 42mg
Bovine serum albumin 18mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds hydroxy tetrahydro pyrimidine and bovine serum albumin then, leaves standstill 1 hour;
2) lyophilizing: a, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 10Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 5 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Tetrahydropyrimidine 26mg
Polyvinylpyrrolidone 10mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds tetrahydropyrimidine and polyvinylpyrrolidone then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 10Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 6 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Tetrahydropyrimidine 58mg
Trehalose 50mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds tetrahydropyrimidine and trehalose then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 12Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃, kept dry 5 hours, promptly get with 0.1 ℃/minute programming rate.
Embodiment 7 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Tetrahydropyrimidine 72mg
Bovine serum albumin 23mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds tetrahydropyrimidine and bovine serum albumin then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 10Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 8 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Tetrahydropyrimidine 60mg
Polyvinylpyrrolidone 3mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds tetrahydropyrimidine and polyvinylpyrrolidone then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 12Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 9 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Tetrahydropyrimidine 15mg
Polyvinylpyrrolidone 12g
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds tetrahydropyrimidine and polyvinylpyrrolidone then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 15Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 10 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Tetrahydropyrimidine 17mg
Polyvinylpyrrolidone 19mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds tetrahydropyrimidine and polyvinylpyrrolidone then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 9Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 11 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Hydroxy tetrahydro pyrimidine 20mg
Polyvinylpyrrolidone 10mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds hydroxy tetrahydro pyrimidine and polyvinylpyrrolidone then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 10Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 12 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Hydroxy tetrahydro pyrimidine 12mg
Bovine serum albumin 15mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds hydroxy tetrahydro pyrimidine and bovine serum albumin then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 10Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 13 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Tetrahydropyrimidine 18mg
Trehalose 59mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds tetrahydropyrimidine and trehalose then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 10Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 14 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Tetrahydropyrimidine 11mg
Trehalose 75mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds tetrahydropyrimidine and trehalose then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 10Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 15 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Tetrahydropyrimidine 11mg
Bovine serum albumin 32mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds tetrahydropyrimidine and bovine serum albumin then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 7Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
Embodiment 16 injection erythrocyte lyophilized powders prescription and preparation
Erythrocyte cell 2 * 10
9Cell/ml 1000ml
Tetrahydropyrimidine 19mg
Trehalose 41mg
1) dosing:
With normal saline washing 2 times, using sterilized water for injection to be made into concentration is 2 * 10 with erythrocyte
9The suspension of cell/ml adds tetrahydropyrimidine and trehalose then, leaves standstill 1 hour;
2) lyophilizing:
A, pre-freeze: the solution that will be sub-packed in cillin bottle is reduced to-50 ℃ with 10 ℃/minute speed from-20 ℃, keeps freezing 2 hours; B, primary drying: be evacuated to 10Pa, be warming up to-10 ℃ with 5 ℃/minute programming rate then, kept dry 6 hours; C, redrying: behind the primary drying, be warming up to 25 ℃ with 0.1 ℃/minute programming rate, kept dry 5 hours, the sterilization packing promptly gets.
The acute toxicity testing of embodiment 17 injection erythrocyte freeze-dried powder preparations
SPF level KM mice; Male and female half and half; Body weight 16g-30g is divided into 17 groups at random after weighing, one group of tail vein injection gives not freeze dried erythrocyte; All the other 16 groups respectively intravenous injection give embodiment 1-embodiment 16 prepared lyophilized powder (lyophilized powder redissolves with normal saline), calculating dosage with the erythrocyte number of cells is 10
12/ only, observed 14 days continuously after the administration.Observation item comprises toxic reaction, body weight and histopathology.The result shows that mouse tail vein injection gives the erythrocyte lyophilized powder, counts 10 with the erythrocyte number of cells
14/ the time, the reaction of no overt toxicity, before and after the administration and experiment mice body weight no significant difference when finishing, pathological study is found significantly unusual.Explain that injection erythrocyte freeze-dried powder preparation of the present invention safety is good.
The long term toxicity test of embodiment 18 injection erythrocyte freeze-dried powder preparations
The Wister rat; Male and female half and half; 420g-600g is divided into 17 groups at random after weighing, one group of tail vein injection gives not freeze dried erythrocyte; All the other 16 groups respectively intravenous injection give embodiment 1 to embodiment 16 prepared lyophilized powder (lyophilized powder redissolves with normal saline), calculating dosage with the erythrocyte number of cells is 2 * 10
9/ only, administration was 1 time in per two days, and totally 30 days, convalescent period was 14 days.During the administration and in convalescent period; Observe before the rat administration and after the administration and carry out gross examination of skeletal muscle; The body weight of rat before the general symptom of record rat, the record administration and after the administration, and regularly carry out hematology, urine and blood biochemical analysis reaches the electrocardiogram of rat after the administration before the record administration.The result shows that each treated animal body weight and matched group do not have significant difference, and gross examination of skeletal muscle is no abnormal.The hematology of experimental group animal, urine and blood physicochemical data and matched group do not have significant difference.The safety that four injection erythrocyte freeze-dried powder preparation long term administrations are described is good, and toleration meets the requirement of clinical application.
The erythrocyte response rate of embodiment 19 injection erythrocyte freeze-dried powder preparations
The erythrocyte in vitro cell response rate=(erythrocyte number after the rehydration/lyophilizing proerythrocyte number) * 100%
Embodiment 1-embodiment 16 prepared lyophilized powders are used the normal saline rehydration, number after reaching rehydration before each group erythrocyte cell rehydration is counted, calculate and respectively organize the erythrocyte cell response rate (get for every group and average after three duplicate samples are measured).The result shows; Each erythrocyte in vitro cell response rate (average) of organizing lyophilized powder is 85.7%-96.0%; Wherein the erythrocyte in vitro cell response rate of the lyophilized powder of being made up of tetrahydropyrimidine or hydroxy tetrahydro pyrimidine and polyvinylpyrrolidone of embodiment 1, embodiment 2, embodiment 9 and embodiment 11 preparations concentrates on 87.1%-98.3%, is significantly higher than other and respectively organizes.
Morphological observation after the erythrocyte rehydration of embodiment 20 injection erythrocyte freeze-dried powder preparations
After the injection erythrocyte lyophilized powder that embodiment 1 to embodiment 16 is prepared redissolved with normal saline, the erythrocyte smear was done Ji's nurse Sa dyeing (Giemsa stain), fresh red blood cell behind smear staining equally as normal control; Observation by light microscope is also taken a picture.After the result shows that embodiment 1 to embodiment 16 prepared injection erythrocyte lyophilized powder redissolves with normal saline, the red cell morphology no abnormality seen.Red cell morphology after the embodiment 9 injection erythrocyte lyophilized powder rehydrations is seen Fig. 1.
Adenosinetriphosphataes and glucose-6-phosphate dehydrogenase (G6PD) determination of activity after the erythrocyte rehydration of embodiment 21 injection erythrocyte freeze-dried powder preparations
The injection erythrocyte lyophilized powder that embodiment 1 to embodiment 16 is prepared uses kit measurement adenosinetriphosphataes (ATP) and glucose-6-phosphate dehydrogenase (G6PD) (G-6-PD) activity after redissolving with normal saline.Fresh red blood cell and 2% standard red cell suspension are done same mensuration as contrast.The result sees table 1.
Table 1
Group |
ATP active (10
7/U)
|
G-6-PD active (%) |
Fresh red blood cell |
17.5±2.8 |
84.2±5.3 |
2% standard red cell suspension |
10.2±5.6 |
59.4±4.7 |
After the embodiment 1 lyophilizing erythrocyte rehydration |
16.9±3.0 |
83.8±7.4 |
After the embodiment 2 lyophilizing erythrocyte rehydrations |
17.2±9.5 |
84.0±6.1 |
After the embodiment 3 lyophilizing erythrocyte rehydrations |
16.6±6.4 |
81.7±3.9 |
After the embodiment 4 lyophilizing erythrocyte rehydrations |
16.0±5.7 |
82.1±9.4 |
After the embodiment 5 lyophilizing erythrocyte rehydrations |
18.3±4.1 |
83.5±1.0 |
After the embodiment 6 lyophilizing erythrocyte rehydrations |
16.6±8.6 |
82.4±6.7 |
After the embodiment 7 lyophilizing erythrocyte rehydrations |
16.2±6.5 |
81.8±3.2 |
After the embodiment 8 lyophilizing erythrocyte rehydrations |
17.5±4.4 |
83.0±2.6 |
After the embodiment 9 lyophilizing erythrocyte rehydrations |
17.1±2.9 |
84.1±6.9 |
After the embodiment 10 lyophilizing erythrocyte rehydrations |
18.1±0.3 |
83.5±2.8 |
After the embodiment 11 lyophilizing erythrocyte rehydrations |
17.6±5.2 |
83.2±5.3 |
After the embodiment 12 lyophilizing erythrocyte rehydrations |
16.8±8.3 |
81.9±6.6 |
After the embodiment 13 lyophilizing erythrocyte rehydrations |
16.1±4.6 |
81.1±9.7 |
After the embodiment 14 lyophilizing erythrocyte rehydrations |
16.6±3.1 |
82.4±3.5 |
After the embodiment 15 lyophilizing erythrocyte rehydrations |
16.0±1.2 |
83.0±4.9 |
After the embodiment 16 lyophilizing erythrocyte rehydrations |
16.7±3.5 |
80.7±5.1 |
The result shows, the erythrocyte lyophilized powder of embodiment 1 to embodiment 16 preparation, and ATP activity and G-6-PD activity do not have significant difference in fresh red blood cell after the rehydration, significantly are superior to red cell suspension commonly used at present.