CN106267418B - Female tire blood group incompatibility antibody adsorbing therapy instrument - Google Patents
Female tire blood group incompatibility antibody adsorbing therapy instrument Download PDFInfo
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- CN106267418B CN106267418B CN201610539276.6A CN201610539276A CN106267418B CN 106267418 B CN106267418 B CN 106267418B CN 201610539276 A CN201610539276 A CN 201610539276A CN 106267418 B CN106267418 B CN 106267418B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
- A61M1/362—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits changing physical properties of target cells by binding them to added particles to facilitate their subsequent separation from other cells, e.g. immunoaffinity
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
- A61M1/3621—Extra-corporeal blood circuits
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
- A61M1/3687—Chemical treatment
- A61M1/3689—Chemical treatment by biological cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M2202/00—Special media to be introduced, removed or treated
- A61M2202/04—Liquids
- A61M2202/0413—Blood
- A61M2202/0429—Red blood cells; Erythrocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M2205/00—General characteristics of the apparatus
- A61M2205/02—General characteristics of the apparatus characterised by a particular materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M2210/00—Anatomical parts of the body
- A61M2210/12—Blood circulatory system
Abstract
The present invention relates to female tire blood group incompatibility antibody adsorbing therapy instrument of medical domain, it is characterized in that with 4 DEG C and 37 DEG C of the positive O-shaped red blood cell of physiological saline cleaning Rh, Washed Red Blood Cells are replaced with the PB lysate of the PH7.4 of 25 and 35mmol/L again, preparation removes the ghost of hemoglobin and the antigen containing D thoroughly, 80% cell concentration is made into the alserver's solution of 3.5% mannitol, assemble cylindrical adsorption device made of high-biocompatibility material, sealing, set 4 DEG C of preservations, sieve is arranged in exit in its absorber, constitute the defence line for preventing cell fragment from filtering out, after blood plasma filtration, Rh antibody is combined into fixed compound by Rh positive red blood cell therein, the red cell debris that is destroyed and it is in combination made of macromolecular complex be adsorbed the sieve detention of device, removal is caused a disease The blood plasma of substance is fed back after filtering out from absorber, to treat blood group incompatibility Hemolysis by removing blood plasma Rh antibody.
Description
Technical field
The present invention relates to female tire blood group incompatibility antibody adsorbing therapy instrument of medical domain, and it is pregnant to be mainly used for female tire blood group incompatibility
The removing of anti-fetal red blood cells antibody and red blood cell lysate in woman's blood plasma.
Background technique
Fetus inherits the gene element of father and each half of mother, and fetal red blood cells can carry the antigen from male parent,
The blood group for showing as fetus is different from parent.After the red blood cell of fetus enters the blood circulation of parent, the immune of parent is induced
System generates antibody, and antibody enters fetal circulation system by placenta, in conjunction with fetal red blood cells, makes fetal red blood cells quilt
It destroys, leads to female tire blood group incompatibility hemolytic disease of fetus an d neonate.
Human erythrocyte's blood group has 26 kinds, including the blood groups such as abo blood group, Rh blood group and MN, Lew, Kell and Fya, but
The blood group of female tire blood group incompatibility Hemolysis can be caused with Rh blood group and abo blood group to be most common.Rh blood group antigens are contaminated by No. 1
The allele of 3 pairs of close linkages determines on colour solid, shares 6 kinds of antigens, i.e. C and c, D and d, E and e.Since D antigen is earliest
It is found, antigenicity is most strong, therefore clinically all D antigen positive persons are known as the Rh positive, and no D antigen person is known as Rh feminine gender.Rh blood
The antigenicity of type antigen determines the severity of Hemolysis, and D antigen can cause serious Hemolysis, is secondly E antigen, then
Secondary is C, c and e antigen, and the antigenicity of d antigen is most weak, and anti-d antibody there is no to find at present.Rh is negative in China's Chinese Han Population
Frequency be about 0.4%, and the frequency of ethnic group of the Northwest Rh feminine gender is up to 10% or more.
Although the incidence that abo blood group does not conform to is very high, fetus haemolysis incidence is very low, even if haemolysis occurs, symptom is also
Relatively light, few that nuclear icterus and oedema occurs, the gestational period is not necessarily to specially treated.And Rh blood group incompatibility is although rare, but it causes mother
The severity extent of tire blood group incompatibility Hemolysis will overweight abo blood group and not conform to, so the diagnosis and prevention to Rh blood group incompatibility are extremely
It is important.Rh blood group incompatibility generates the Rh antibody (the anti-D of IgG) of a large amount of anti-fetal red blood cells due to parent, into fetus body in after
Destroy a large amount of fetal red blood cells, make fetus severe anemia, anoxic, heart failure, hepar damnification, Hypoproteinemia, anasarca,
Hydrothorax, ascites even Intrauterine Fetal Death.In non-neonate, Rh blood group incompatibility haemolysis does not conform to compared with abo blood group there is the jaundice time
It is early, occur after birth in 12 hours earliest, majority appeared in 24 hours.Since a large amount of bilirubin that haemolysis generates cannot
It is excluded in time from liver, aggravates icterus neonatorum, a large amount of bilirubin penetrates into brain cell and causes nuclear icterus, often dies of tight
Weight anaemia, heart failure, nuclear icterus, the death rate are very high.
Although Rh antibody, which enters the rear red blood cell by destroying fetus in fetus body, can cause serious Hemolysis, certain
A little pregnant woman also can be by injecting Rh antibody in advance come Rh (D) positive red blood cell sensitization body of pre- anti-intrusion, and then can prevent Rh
The generation of Hemolysis.Since current most countries have forbidden being carried out that D antigen is immune with volunteer, the polyclonal anti-D's of Ig comes
Source is extremely limited, and utilizes the anti-D of Epstein-Barr virus conversion human B lymphocyte secretion Ig or Epstein-Barr virus conversion combination cell fusion system
The standby anti-D of Ig, there are still very by the anti-D of Ig secreted by the bone-marrow-derived lymphocyte strain or hybridoma cell strain that use EBV conversion in vivo
More problems, such as contained EBV have potential pathogenic；It may be with pathogen such as HIV and hepatitis virus；Use hybridoma technology
The anti-D albumen containing source of mouse of the Ig of production easily causes allergic reaction or the oncogene with mouse, so by directly in vivo
The injection anti-D of Ig is just restricted to prevent Rh Hemolysis.
Clinically mainly there are pregnancy period plasma exchange, fetal transfusion, termination to the treatment of female tire blood group incompatibility Hemolysis at present
Gestation and neonatal exchange transfusion treatment.Anti- D potency needs to consider intrauterine transfusion 32 or more, and fetal transfusion includes that fetus is intraperitoneal defeated
Blood and the intravascular blood transfusion of fetus, all have certain risk, and none is proved effectively；Hyperbilirubinemia of newborn person can adopt
With drug therapies such as blue light illumination, intravenous injection of immunoglobulin, Blood exchanging therapy, Blood exchanging therapy are still that treatment is newborn when necessary
The main method of youngster's Hemolysis；Anti- D potency need to consider replacement therapy of blood plasma, pregnant woman blood plasma displacement 64 or more during gestation
It is exactly the haemocyte and blood plasma for filtering method separation pregnant woman in circulation path in vitro, removes blood plasma and the displacement liquid with equivalent
(fresh frozen plasma or 5% albumin) supplement feed back, each 2 000~3 000ml of replacement amount, 20~30ml/min of speed,
2~4h for the treatment of time, every 1~3d are treated 1 time；Or pregnant woman's whole blood about 400ml is taken every time, its blood plasma is removed about after low-temperature centrifugation
300ml supplements equivalent homotype fresh plasma, also defeated Autoerythrocyte (RBC).Plasma exchange can with the plasma volume that is removed at
The titre (potency) of pathogenic antibody is reduced, ratio so as to extend fetus in female intracorporal survival and growth and development time, energy
The time for postponing terminal pregnancy, be female tire ABO, Rh or other blood group incompatibilities pregnant woman prevent miscarriage in clinical early stage it is good in treatment
Good selection, tool has a better effect, also without other adverse reactions.But plasma exchange can only remove the partial antibody in blood, no
It can stop antibody to continue to generate, the female tire blood group incompatibility Hemolysis occurred can not be reversed, need to carry out blood incessantly
Slurry displacement is just effective, is only applicable to that the pregnant woman of fetus edema once occurred before gestation 20~22 weeks or spouse is pathogenic antigens
Homozygote person, especially while removing part pathogenic antibody, also eliminate together a large amount of blood plasma (multiple beneficial at
Part), although being supplemented with displacement liquid, it can not supply completely and be removed blood plasma and its various beneficials, and substitute
Replace that liquid measure is big, somewhat expensive, supplement allosome blood plasma easily causes the various side effects such as infectious disease and infusion reaction, which limits
The generally development of plasma exchange.
So there is an urgent need to a kind of only removal pathogenic antibody, not removing blood plasma and its multiple beneficial composition, without using blood
Starch the treatment new method of female tire blood group incompatibility Hemolysis cheap, safe, without side-effects of displacement liquid.
Summary of the invention
In order to solve the problems, such as to attack the treatment for the female tire blood group incompatibility Hemolysis being unable to long, present inventors have proposed the present invention.
The invention aims to provide female tire blood group incompatibility antibody adsorbing therapy instrument；Another object is to provide for absorber
Preparation and application method.
The object of the present invention is achieved like this: the positive O-shaped red blood cell of the fresh Rh of sufficient amount is taken, respectively at 4 DEG C and 37
With physiological saline cleaning 4 times under conditions of DEG C, to remove erythrocyte surface immune antibody that may be present or natural antibody,
Then with 0.04mol/L Na2HPO4With 0.04mol/L NaH2PO4The osmotic concentration of preparation is the PH7.4 of 25 and 35mmol/L
PB lysate replace Washed Red Blood Cells, finally with physiological saline cleaning until supernatant it is colourless, in red blood cell of dialysing to people
The harmful hemoglobin of body is centrifuged 10min through 4 DEG C of 2500r/min, and gained cell precipitation is the ghost of the antigen containing D,
After detection cellular morphology and antigenicity, cylindrical adsorption device made of high-biocompatibility material is assembled, until 4/5, it is added
3.5% mannitol alserver's solution makes red blood cell concentration up to 80%, and jog mixes, and 4 DEG C of preservations are set in sealing, absorber
Top and the bottom are respectively provided with cell screen clothes, and top diameter is 500 mesh, and bottom diameter is 50 mesh, and 200 aim cell strainers are arranged in liquid outlet, constitute
It prevents cell fragment from entering the second defence line of circulation, buffer area is set between liquid entrance and mesh screen so that extracorporal circulatory system
Stabilization, when blood plasma flows through absorber, Rh antibody is combined into immune complex by ghost therein, and what is be destroyed is red
Cell fragment and it is in combination made of macromolecular immune complex be adsorbed the sieve detention of device, remove the blood of morbid substance
Slurry is fed back in vivo after filtering out from absorber, removes blood plasma Rh antibody and red blood cell lysate with this.
Rh antibody is the virulence factor of female tire Rh blood group incompatibility Hemolysis, and Rh positive red blood cell is the natural anti-of Rh antibody
Original, can in conjunction with Rh antibody and by absorption remove, the present invention chooses the positive O-shaped red blood cell of Rh, is made with the dialysis of PB lysate
Removal is harmful to the hemoglobin of human body but retains natural antibody of the antigenicity of absorption Rh antibody without adsorbing anti-A and anti-B
Ghost, being followed by 3.5% mannitol alserver's solution is that medium assembles absorber, is easy to periodically save, Rh is positive
Red blood cell is easy to get, and preparation method is easy, and because the special sieve in absorber exit is anti-to red cell debris and macromolecular antigen
The barrier properties of nanocrystal composition, form Rh positive red blood cell absorption Rh antibody and absorber mesh screen machinery detention red blood cell is molten
The double barrier for solving product, flows through the red blood cell lysate quilt of the Rh antibody and haemolysis patient in the blood plasma of absorber
Absorption is removed, and purified blood plasma is fed back, can be with rhesus monkey erythrocytes substitute mankind Rh positive red blood cell and prepare Rh blood group not
The treatment cell for closing Hemolysis, is a kind of saving mankind's blood source, only removes pathogenic antibody and red blood cell dissolved matter, does not remove
Blood plasma and its multiple beneficial composition, female tire blood group incompatibility Hemolysis cheap, safe, without side-effects without plasma exchange liquid
Treatment new method.
Specific embodiment
Fig. 1 is the application schematic diagram of the female tire blood group incompatibility antibody adsorbing therapy instrument proposed according to the present invention.
Fig. 2 is the schematic diagram of internal structure of the plasma separator proposed according to the present invention.
Fig. 3 is the schematic diagram of internal structure of the absorber proposed according to the present invention
In Fig. 1, one end of arterial blood line pipe (1) is connected with arteries, and the other end is through heparin pump (2) and blood pump
(3) it is connected with plasma separator (4), plasma separator (4) absorption in parallel with 2 through blood plasma pump (6) and circulation line (7)
Device (8), absorber (9) be connected, be then successively connected with circulation line (10), venous line (5), venous line (5) it is another
End is connected with vein blood vessel.
In Fig. 2,1 is plasma separator, and 2 be plasma separator inner cavity, and 3 be the micropore on the tube wall of plasma separator inner cavity, 4
Being cannot be by the haemocyte of micropore (3), and 5 be the small molecule blood plasma components that can pass through micropore (3), and 6 be outside plasma separator
Chamber, 7 be blood plasma outflux, and 8 be switchable valve.
In Fig. 3,1 is absorber, and 2 be the Rh positive ghost in absorber, 3 be into absorber Rh it is anti-
Body, 4 be the antigenantibody complex that Rh antibody is formed by Rh positive blood shadow cell combination, and 5 be RBC fragment.
Below with reference to Fig. 1, Fig. 2 and Fig. 3, to the embodiment party of female tire blood group incompatibility antibody adsorbing therapy instrument proposed by the present invention
Case is explained in detail.
One, the preparation of Rh positive ghost
1, the acquisition of Rh positive red blood cell
According to treatment and raw material sources difficulty or ease, cell below is chosen:
(1) positive " O " the type red blood cell of fresh concentration Rh (D) is bought from Zhejiang Province blood station；(2) take newborn in vitro
Discarded umbilical cord Rh (D) positive " O " type erythrocyte；(3) rhesus macaque " O " type red blood cell.(4) containing blood group antigens needed for treating
Cell.
2, main agents
(1) 30mmol/L PB buffer: solution A is 0.04mol/L Na2HPO4(Na2HPO4.12H2O 14.328g dissolution
In 1000ml deionized water, room temperature storage is spare)；Second liquid is 0.04mol/L NaH2PO4(NaH2PO4.2H2O 6.24g is molten
In 1000ml deionized water, room temperature storage is spare).Take respectively solution A 81.0ml mixed with second liquid 19.0ml 40mmol/L
PB, then on the basis of 40mmol/L PB buffer plus deionized water dilutes 0.75 times to become 30mmol/L PB slow
Fliud flushing is made into 25 and 35mmol/L with method in proportion.
(2) anti-D standard items and Rh (D) normal erythrocytes are purchased from Guangzhou Lian Tai Bioisystech Co., Ltd.
The formula of (3) 3.5% mannitol alserver's solutions are as follows: take mannitol 17.5g, citric acid trisodium 1.5g, citron
Sour 0.2g, glucose 7.93g, sodium dihydrogen phosphate 0.94g, adenine 0.14g, sodium chloride 4.97g, are dissolved in the distilled water of 500ml
In.Wherein mannitol dilutes Red Blood Cells Concentrate viscosity, increases cell membrane stability, prevents haemolysis.Sodium citrate and blood or
Ca2+ in blood plasma combines the soluble calcium citrate for being formed and being difficult to dissociate, and reduces Ca2+ in blood, to inhibit blood coagulation
Process generates anticoagulation, and has protective effect to red blood cell, can prevent the generation of haemolysis.Citric acid prevents red blood cell from adding
Add glucose coking in agent, forms buffering pair with sodium citrate, adjust and stablizing solution pH.Glucose is erythrocyte metabolism
Main energy sources, under normal circumstances, the 90% of glucose are generated by anerobic glycolysis, 10% by pentose phosphate pathway
ATP provides red blood cell energy and maintains its service life.Activity level of the blood ATP during 4 DEG C of storages can be improved in adenine.It is red thin
Born of the same parents be to the needs of adenine it is special, adenine can be changed into adenosine phosphate (AMP) by it, and further phosphoric acid metaplasia
At ATP, energy-rich compound source is provided for red blood cell metabolism vigor, greatly prolongs holding time of the blood at 4 DEG C.Chlorine
Changing sodium to adjust solution osmotic pressure is isotonic solution, provides appropriate sodium ion for red blood cell.Sodium dihydrogen phosphate prevents red blood cell poly-
Collection, provides phosphate for energy metabolism of erythrocyte, slows down 2,3-DPG decrease speed.
3, preparation method
(1) the positive O-shaped red blood cell of the fresh Rh of sufficient amount is taken, respectively under conditions of 4 DEG C and 37 DEG C, with 1500r/min
The centrifugal speed of × 5min is cleaned red blood cell 4~5 times with isometric physiological saline, may be adhered to removing erythrocyte surface
Abo blood group and Rh blood group and other immune antibody or natural antibody.
(2) with 0.04mol/L Na2HPO4With 0.04mol/L NaH2PO4The osmotic concentration of preparation is 25 and 35mmol/L
PH7.4 PB lysate, under 4 DEG C and 2500r/min × 10min of centrifugal condition, alternately and repeatedly Washed Red Blood Cells lead to
The alternating variation for crossing osmotic concentration, promotes hemoglobin to give outside red blood cell and be removed.
(3) finally with physiological saline cleaning until supernatant is colourless, with the blood being harmful to the human body in red blood cell of dialysing completely
Lactoferrin, precipitating are the ghost of the antigen containing D.
(4) detection of antigenicity: 1. by ghost and 37 DEG C of incubation 5min of anti-D standard items, the detection of centrifuging and taking supernatant is anti-
- D standard items potency reduces numerical value to determine the antigenicity of ghost.Specific method is: take 10, test tube, respectively number 1~
10, the 1st pipe is added ghost and precipitates 1.0ml, then other each pipes plus physiological saline 0.5ml inhale ghost from the 1st pipe
Precipitating 0.5ml is added to the 2nd pipe, and 0.5ml to the 3rd is inhaled after mixing and is managed, and is diluted to the 10th pipe with same operation, finally pipe is sucked out
0.5ml is discarded, and every pipe adds anti-D standard items 0.5ml, 37 DEG C of incubation 5min to mix gently after centrifugation, to occur being aggregated completely,
The antigenicity of ghost is represented almost without the maximum dilution multiple of free cell, extension rate is bigger, and antigenicity is stronger, inhales
The ability of attached Rh antibody is stronger.Supernatant antibody titer can also be measured with Rh (D) normal erythrocytes to determine ghost
Antigenicity；2. measuring supernatant antibody titer with Rh (D) normal erythrocytes, anti-D standard items potency (known) subtracts supernatant
Antibody titer is the antigenicity (potency) of ghost.Supernatant antibody titer detection method are as follows: take 10 test tubes, respectively
Physiological saline 1ml is added in number 1~10, each pipe, takes supernatant 1ml to be added to removal 1ml after the 1st pipe mixes and goes to the 2nd pipe,
It is diluted to the 10th pipe with same operation, finally pipe is sucked out 1ml liquid and discards, and serum 40 μ l and Rh (D) standard diluted is red
10 μ l of cell mixing, is incubated for 10 minutes, is centrifuged 5 minutes interpretation results, and the agglutination pipe of maximum dilution multiple is antigenicity (effect
Valence), potency should be 1: 1500 or more.
(5) content of hemoglobin detects: with hypotonic destruction ghost, with conventional chemical luminescence analysis or fluorescence analysis
Method, the qualitative content of hemoglobin in quantitative detection ghost should be lower than the reference value lower bound of human normal plasma.
(6) ghost form: observing the form and integrality of ghost under the microscope, should be in circle shadow, no-reflection.
(7) for cellular morphology in circle shadow, no-reflection, highly sensitive routine hemoglobin qualitative detection is feminine gender, quantitative
Detection is lower than the reference value lower bound of human normal plasma, adsorbs the ghost of 1: 1500 or more Rh antibody titer, leaves and takes spare.
Two, the preparation of absorber
1, preparation principle
It is because the free Rh antibody in pregnant woman blood plasma enters fetal blood the present invention is based on female tire Rh blood group incompatibility Hemolysis
It destroys caused by fetal red blood cells, and Rh antibody can be combined absorption, and absorption by corresponding native antigen, that is, Rh positive red blood cell
The shell of device can be made into mechanism preparation of the sieve for only allowing specific small molecule to pass through to stop bigger molecule to pass through.
2, material is prepared
It selects and the close high-biocompatibility material of Human vascular endothelial, it is desirable that good biocompatibility, no complement activation,
No inflammation reaction, the change without leucocyte, blood platelet, blood oxygen pressure, complement C 3 C5a.It is required that passing through covalent, grafting, polymerization
The methods of improve the influence to blood coagulation and oxidative stress of uniformity, hydrophily, reduction of material surface.In absorber inner surface
Add hydrophilic gel, such as solidifies 2 methylacryoyloxyethyl phosphocholines-butyl methacrylate in cellulose acetate film, pass through
It controls wet-spinning procedure and generates CA/PMB30, CA/PMB80 and CA/PMB30-80, biocompatibility can be improved.It will be certain anti-
Condensate matter is solidificated in carrier or absorber inner surface, can inhibit blood clotting, reduces heparin dosage even realization no-rod tractor,
Such as heparin is aggregated on polyacrylonitrile-polyethyleneimine film, the allergic reaction of allergic constitution can be reduced；By heparin covalent knot
Polyether sulfone surface is closed, the mechanical property of polyether sulfone can be kept and improves the anticoagulation function of absorber inner surface.In acetic acid fibre
Covalent immobilisation linoleic acid film on film is tieed up, or the linoleic acid for being covalently bound to polyacrylic acid is grafted onto polysulfones film surface, is ok
There are better histocompatbility and anticoagulant effect.
3, the specification of adsorber enclosure
The hydrostatic column that the bottom diameter for being made 50mm × 60mm is small, top diameter is big, or rectangular, infundibulate is made, volume about 200
~300ml, top and the bottom are equipped with cell screen clothes, and top diameter sieve mesh number is 500 mesh, and bottom diameter sieve mesh number is 50 mesh, liquid outlet
Place's setting mesh number is 200 aim cell strainers, constitutes the second defence line for preventing cell fragment from entering circulation, liquid entrance
It is equipped with buffer area between mesh screen, is conducive to the stability of system circulation.
4, the filling of adherent cell
Rh positive blood shadow cell precipitation prepared by the present invention is taken, absorber is packed into, until 4/5, the red of 3.5% mannitol is added
Cell-preservation liquid makes red blood cell concentration up to 80%, and jog mixes, and capping is set 4 DEG C and saved backup.
Three, the preparation of plasma separator
1, it preparation principle: is prepared according to the molecular size of haemocyte and blood plasma components.Such as visible component (blood in blood of human body
Cell) size are as follows: normocyte is about 7 microns (μm), is the discoid cell of concave-concave；Leucocyte is divided into 5 kinds, in
Property about 12 μm of granulocyte, eosinophil is more bigger, and basophilic granulocyte and neutrophil leucocyte are close, small lymphocyte 6-8
μm, approximate with red blood cell, monocyte is maximum, and about 15-20 μm.Blood platelet is disc, 1~4 micron to 7~8 microns of diameter
It differs, the platelet mean diameter of people is 2-4 microns, 0.5~1.5 micron thick.
2 prepare material: poly-vinegar non-woven fabrics, acetate fiber, absorbent cotton etc. can be selected, it is desirable that good biocompatibility, hardly
Activating complement, the change for not causing inflammatory reaction, not causing leucocyte, blood platelet, blood oxygen pressure, complement C 3, C5a.It can lead to
Cross covalently, grafting, the methods of polymerization improve the structure of material, adjust the microinhomogeneities on surface, hydrophily, reduction to solidifying
The influence of blood and oxidative stress, the generation to improve sieving adequacy and biocompatibility, reduction complication.
3 specifications: for the shape of separator, cylindricality knot can be prepared into as filter core with materials such as acetate fiber or absorbent cotton
Structure is prepared into the shapes such as flat structure as filter core with materials such as poly-vinegar non-woven fabrics；By haemocyte and blood plasma components to be separated
Molecular size determines aperture.Plasma separator according to the present invention property stabilization, good biocompatibility, permeability are high
Hollow fibre type filter is made in high molecular polymer, and hollow-fiber film diameter is 270~370 μm, and film thickness is 50 μm, aperture
It is 0.2~0.6 μm, fibre length is 13.5~26 μm.Blood plasma filtration is only permitted in the hole, but can stop all cell components.
Four, application of the invention
Extracorporal circulatory system branch need to be collectively constituted with associated components.
1, the component and purposes of extracorporal circulatory system branch
(1) absorber: the positive ghost of internal Rh (D) is for removing Rh antibody；The net of adsorber enclosure inlet and outlet
Sifter device plays the role of filtering out the macromolecular complex that RBC fragment, Rh antibody and RBC fragment are formed.
(2) washed corpuscles and blood plasma plasma separator: are used for.
(3) sound pulse pressure monitors: in addition the main stopping state to dynamic monitoring absorber micropore of arterial blood pressure monitoring is used
To monitor extracorporal circulatory system thrombus, solidification and the variation of pressure.When blood flow deficiency, angiosthenia will be reduced；When having blood coagulation, blood
Bolt is formed, especially absorber blockage of the micro orifice when, angiosthenia will increase；Vein pressure monitoring is used to monitor pipeline blood reflux
Pressure, when absorber blockage of the micro orifice, blood coagulation, thrombosis, blood flow is insufficient and venous return syringe needle falls off when, vein
Pressure will decline, if bloody path return pipe distortion blocking or reflux syringe needle block, vein pressure will be increased.
(4) air monitering (Air Detector): the air bubble for monitoring blood pathway generally uses ultrasonic listening
Principle, in order to avoid patient occur air embolism and be arranged.When having monitored air bubble, detection system can drive it is dynamic,
Vein bloody path folder carrys out blocking blood flow, prevents dangerous generation.
(5) blood pump (Blood Pump): for pushing blood circulation going on smoothly with maintenance therapy, usual blood pump part
Often there is rotary test speed function, to monitor the blood circumstance of patient, therefore blood pump runner and the setting of groove spacing are accurate,
And often adjustment is needed, and the case where according to bloody path pump line, spacing is generally set as 3.2~3.3mm, can not be too loose, otherwise can
Cause blood flow detection inaccurate；Also can not be too tight, it otherwise will cause pipe breakage.
(6) heparin pump (Heparin Pump): heparin pump is equivalent to the micro-injection pump clinically applied, to continue to
Injecting heparin in sieving pipeline (patient blood) contacts with air since the blood of patient recycles in vitro, is easy to happen blood coagulation
Phenomenon anticoagulative can be occurred using heparin pump.
It additionally include temperature control system, liquid mixing system, off gas system, monitored conductivity system, ultrafiltration monitoring and leakage
The parts such as blood monitoring.In short, being expected to be further development of automation, human nature on the basis of crucial constitution system of the invention
Change, personalization, modularization, automatic monitoring and regulation, voluntarily judge the micro computers such as alarm reason and ring off signal at liquid crystal display
Processing system.
2, application method of the invention
(1) it installs: with each portions such as sterile working connecting components, including plasma separator, absorber and each circulation line.
(2) it is vented: with sterile saline filling liquid separator, absorber and each circulation line, excluding separator, absorber
And its gas, bubble in circulating line, it goes through, confirmation after gas, bubble without using.
(3) lead to liquid: arterial blood line pipe 1 being connected into arteries, goes through again be vented whether complete, liquid in operation
Whether stream is unobstructed, and flow liquid in pipe is avoided to pollute.
(4) anticoagulant: to be injected from heparin pump into liquid stream anti-coagulants (heparin), be for the first time 2500U or 20~30U/kg.
(5) start: arterial blood line pipe (1) is connected to the arteries of curer, venous line (5) are connected curer's
Then vein blood vessel opens blood pump, blood flow is 100~150ml/min, such as Fig. 1, when arterial blood is through arterial blood line pipe
(1) enter plasma separator (4), the blood plasma separated is reached through circulation line (7) under the action of blood plasma pump (6) and adsorbed
Device (8) begins releasing blood plasma wait be full of blood plasma, about 10 minutes, flows out through circulation line (10), synchronous to fill to absorber (9)
Note blood plasma starts again at perfusion blood plasma when the blood plasma in absorber (8) has nearly flowed, and absorber (9) is begun releasing at this time
Blood plasma, two absorbers (8) in parallel, absorbers (9) are alternately.Such as Fig. 2, separated when blood to be separated enters blood plasma
When inner cavity (2) of device (1), the effect through valve (8) can be entered by the small molecule blood plasma and its composition (5) of micropore (3) and be divided
Exocoel (6) from device is then flowed out through plasma outlet port (7), and cannot be flowed by the haemocyte (4) of micropore (3) through valve (8)
Out.Such as Fig. 3, when Rh antibody (3) enters absorber (1) with blood plasma, the Rh positive ghost (2) being adsorbed in device is combined
No longer move down at antigen antibody complex (4), the red cell debris (5) that is destroyed and it is in combination made of divide greatly
Sub- antigen antibody complex cannot be by the mesh screen that absorber exports by detention, purified blood plasma and plasma separator point
It separates out after the haemocyte come converges and feeds back, so until knot is just declared in the plasma circulation amount (usually 9L) being previously set, treatment
Beam, if mating computer program controls, entire therapeutic process is controlled by computer, and can detect working condition at any time, uses meeting
More convenient, automation and safety.
Five, the verifying of practical application of the present invention
In order to verify the effect of absorber removes Rh antibody, the present invention devises easy test method: taking the 2.5 of sterilizing
It is appropriate followed by drawing to 250mm scale to draw Rh (D) positive " O " type erythroprecipitin for × 300mm Westergren's blood sedimentation tube 9
The heat preservation after 100 DEG C dissolve in 42 DEG C of 1.0% spare agarose C1-4B, become semisolid after agarose is cooling, can general
Red blood cell is fixed in pipe, and easy garden cylindricality absorber is made with this.In addition the Fresh Frozen of Zhejiang Province center blood station is bought
Blood plasma 200mL and Rh antibody (the anti-D type serum dry powder standard items of people, Guangzhou Lian Tai Bioisystech Co., Ltd), with fresh plasma
It is made into 1: 200,1: 300,1: 600 antibody titer, routinely Rh (anti-D) titre detection method (reference book), further examined
It surveys and confirms whether above-mentioned prepared antibody titer is consistent, then (Rh) antibody (titre) before referred to as filtering respectively takes 10ml filter preceding anti-
Body is injected separately into the upper end blank pipe of 3 blood sedimentation tubes, after flowing through the red blood cell in blood sedimentation tube, efflux is collected, after referred to as filtering
(Rh) antibody, routinely Rh (anti-D) titre detection method confirms titre, then respectively by antibody after filter through positive red containing Rh (D)
Make to filter out for the 2nd time in the blood sedimentation tube of cell, is so repeated 3 times filtration and antibody titer detection, as a result (table 1) illustrates, Rh antibody
After filtering easy absorber, most of Rh antibody by corresponding Rh (D) positive red blood cell adsorb, through the 1st time, the 2nd time,
3rd time filtration after, the average titer of Rh antibody respectively from 1: 367 before filter be reduced to filter after 1: 183,1: 58,1: 29, explanation
With the increase of filtration number, Rh antibody can be adsorbed constantly by Rh (D) positive red blood cell
Titre testing result (1/x) before and after cleanser of 1 Rh antibody of the table filtration containing ghost
Claims (7)
1. a kind of female tire blood group incompatibility antibody adsorbing therapy instrument for medical domain, which is characterized in that the therapeutic equipment is served as reasons
The vitro Adsorption device that plasma separator and absorber are constituted, the plasma separator is used for washed corpuscles and blood plasma, described
Cell screen clothes are arranged in absorber, for stopping red blood cell dissolved matter, are perfused inside the absorber to contain 3.5% mannitol
The cell suspension for the 80% ghost concentration that alserver's solution is made into, it is anti-for adsorbing the Rh flowed through in the blood plasma of absorber
Body, the PB lysate through 25mmol/L and 35mmol/L alternately washs Rh positive cell and prepares the ghost respectively.
2. mother's tire blood group incompatibility antibody adsorbing therapy instrument according to claim 1, which is characterized in that contained with 81.0ml
0.04mol/L Na2HPO4Solution A and 19.0ml NaH containing 0.04mol/L2PO4Second liquid be made into after 40mmol/L with deionization
Water is diluted to the PB lysate of the PH7.4 of 25mmol/L and 35mmol/L.
3. mother's tire blood group incompatibility antibody adsorbing therapy instrument according to claim 1, which is characterized in that 3.5% sweet dew
The alserver's solution of alcohol is mannitol 17.5g, citric acid trisodium 1.5g, citric acid 0.2g, glucose 7.93g, biphosphate
Sodium 0.94g, adenine 0.14g, sodium chloride 4.97g, are dissolved in the distilled water of 500ml.
4. mother's tire blood group incompatibility antibody adsorbing therapy instrument according to claim 1, which is characterized in that the Rh positive cell
Including the mankind and positive " O " the type red blood cell of rhesus macaque Rh.
5. mother's tire blood group incompatibility antibody adsorbing therapy instrument according to claim 1, which is characterized in that outside the absorber
Shell volume is 200~300ml, and entrance top diameter cell screen clothes mesh number is 500 mesh, and exit bottom diameter cell screen clothes mesh number is 50
Mesh.
6. mother's tire blood group incompatibility antibody adsorbing therapy instrument according to claim 1, which is characterized in that the plasma separator
Hollow fibre filter membrane diameter is 270~370 μm, and film thickness is 50 μm, and aperture is 0.2~0.6 μm, and fibre length is 13.5~26
μm, blood plasma can be filtered but all cell components cannot be filtered.
7. the preparation method of adherent cell, special in mother's tire blood group incompatibility antibody adsorbing therapy instrument according to claim 1
Sign is that the cell of antigen needed for treating Hemolysis containing blood group incompatibility is cleaned with 4 DEG C and 37 DEG C of physiological saline respectively, gone
Immune antibody or natural antibody except erythrocyte surface attachment, the PH7.4 for being then 25 and 35mmol/L with osmotic concentration
PB lysate alternately washs cell, with the hemoglobin in red blood cell of dialysing, prepares the ghost of the antigen containing D, detects cell
Form and antigenicity assemble cylindrical absorber, the alserver's solution of 3.5% mannitol are added, reaches red blood cell concentration
80%, jog mixes, and capping is set 4 DEG C and saved backup.
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| CN201610539276.6A CN106267418B (en) | 2016-07-01 | 2016-07-01 | Female tire blood group incompatibility antibody adsorbing therapy instrument |
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| CN109157694A (en) * | 2018-07-01 | 2019-01-08 | 翁炳焕 | A kind of mother's tire blood group incompatibility immunoadsorption therapy instrument |
| CN109157695A (en) * | 2018-07-19 | 2019-01-08 | 翁炳焕 | Based on the female tire blood group incompatibility therapeutic device for removing pathogenic antibody |
| CN111537315B (en) * | 2020-07-13 | 2020-10-02 | 天津德祥生物技术有限公司 | Erythrocyte membrane separation liquid and erythrocyte membrane separation method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3975156A (en) * | 1975-11-10 | 1976-08-17 | Ortho Diagnostics, Inc. | Method and material for detecting and quantitating fetal erythrocytes in adults |
| CN102264410A (en) * | 2008-12-01 | 2011-11-30 | 通用电气公司 | System and method for separating cells from body fluids |
| CN104818280A (en) * | 2015-04-24 | 2015-08-05 | 深圳市血液中心 | Single-stranded aptamer and application thereof |
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| US9182389B2 (en) * | 2011-02-22 | 2015-11-10 | Chrome Red Technologies, Llc | Detection of specific antigens in a population of antigens |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3975156A (en) * | 1975-11-10 | 1976-08-17 | Ortho Diagnostics, Inc. | Method and material for detecting and quantitating fetal erythrocytes in adults |
| CN102264410A (en) * | 2008-12-01 | 2011-11-30 | 通用电气公司 | System and method for separating cells from body fluids |
| CN104818280A (en) * | 2015-04-24 | 2015-08-05 | 深圳市血液中心 | Single-stranded aptamer and application thereof |
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Effective date of registration: 20200821 Address after: 310006 No. 1, bachelor Road, Zhejiang, Hangzhou Co-patentee after: Shu Lan (Hangzhou) Hospital Ltd. Patentee after: WOMEN'S HOSPITAL, SCHOOL OF MEDICINE, ZHEJIANG University Address before: The 317300 Ring Road Zhejiang County of Xianju province to the north, Xianju County Hospital Patentee before: Weng Binghuan |
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