CN106267421A - Female fetal blood type does not conforms to blood plasma depurator - Google Patents
Female fetal blood type does not conforms to blood plasma depurator Download PDFInfo
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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
Abstract
nullThe female fetal blood type that the present invention relates to medical domain does not conforms to blood plasma depurator，It is characterized in that cleaning fresh Rh positive O type erythrocyte at 4 DEG C and 37 DEG C with normal saline，Washed Red Blood Cells are replaced again with the PB lysate of the PH7.4 of 25 and 35mmol/L，Until upper strata is colourless，Completely harmful in dialysis erythrocyte hemoglobin，The preparation ghost containing D antigen，Assemble in the depurator that high-biocompatibility material is made after detection antigenicity，Repeated high speed centrifugation makes ghost form the lymphocyte depletion agent of highly dense compression，Import and export at depurator and be respectively provided with screen cloth，Constitute the defence line preventing cell debris from leaching，After blood plasma filters，Rh antibody is combined into fixing complex by ghost therein，Destroyed bib and macromolecular complex in combination are cleaned the screen cloth detention of device，The blood plasma removing morbid substance feeds back after depurator leaches，To treat blood group incompatibility Hemolysis by removing blood plasma Rh antibody.
Description
Technical field
The female fetal blood type that the present invention relates to medical domain does not conforms to blood plasma depurator, is mainly used in female tire blood group incompatibility blood of pregnant women
Anti-FE antibody and the removing of erythrocytolysis product in slurry.
Background technology
Fetus inherits father and the gene element of each half of mother, and FE can carry the antigen from male parent,
The blood group showing as fetus is different from parent.After the erythrocyte of fetus enters the blood circulation of parent, the immunity of induction parent
System produces antibody, and antibody enters fetal circulation system through Placenta Hominis, in conjunction with FE, makes FE be broken
Bad, cause female tire blood group incompatibility hemolytic disease of fetus an d neonate.
Human erythrocyte's blood group has 26 kinds, the blood group such as including abo blood group, Rh blood group and MN, Lew, Kell and Fya, but
The blood group that can cause female tire blood group incompatibility Hemolysis is most common with Rh blood group and abo blood group.Rh blood group antigen are by No. 1 dyeing
On body, closely linked allele is determined by 3, has 6 kinds of antigens, i.e. C and c, D and d, E and e.Due to D antigen the earliest by
Finding, antigenicity is the strongest, therefore the most every D antigen positive person is referred to as the Rh positive, is referred to as Rh without D antigen person negative.Rh blood group
The antigenicity of antigen determines the order of severity of Hemolysis, and D antigen can cause serious Hemolysis, is secondly E antigen, is again
C, c and e antigen, the antigenicity of d antigen is the most weak, there is no anti-d antibody at present and finds.Frequency negative for Rh in China's Chinese Han Population
It is about 0.4%, and the frequency of the Northwest ethnic groups Rh feminine gender is up to more than 10%.
Although the incidence rate of ABO incompatibility is the highest, but fetus haemolysis incidence rate is the lowest, even if there is haemolysis, symptom is also
Relatively light, few generation bilirubin encephalopathy and edema, trimester of pregnancy is without special handling.Although and Rh blood group incompatibility is rare, but it causes mother
The severity extent ABO incompatibility to be overweighted of tire blood group incompatibility Hemolysis, so to the diagnosis of Rh blood group incompatibility and prevention extremely
Important.Rh blood group incompatibility produces the Rh antibody (the anti-D of IgG) of a large amount of anti-FEs due to parent, rear broken in entering fetus body
Bad substantial amounts of FE, makes fetus severe anemia, anoxia, heart failure, hepar damnification, hypoproteinemia, hydrosarca, breast
Water, ascites even Intrauterine Fetal Death.At non-neonate, there is the jaundice time early in Rh blood group incompatibility haemolysis relatively ABO incompatibility,
Early occur that after birth in 12 hours, majority occurred in 24 hours.Due to haemolysis produce a large amount of bilirubin can not in time from
Liver get rid of, make neonatal jaundice increase the weight of, substantial amounts of bilirubin penetrate into brain cell and cause bilirubin encephalopathy, often die from severe anemia,
Heart failure, bilirubin encephalopathy, mortality rate is the highest.
Although serious Hemolysis can be caused by destroying the erythrocyte of fetus after in Rh antibody entrance fetus body, but certain
A little anemia of pregnant woman also can carry out Rh (D) the positive red blood cell sensitization body of pre-anti-intrusion by injecting Rh antibody in advance, and then can prevent Rh
The generation of Hemolysis.Owing to current most countries has forbidden carrying out D antigen immune, the source of the anti-D of polyclone Ig with volunteer
The most extremely limited, and utilize the Epstein-Barr virus conversion human B lymphocyte secretion anti-D of Ig or Epstein-Barr virus to convert and combine cell fusion preparation
The anti-D of Ig, the anti-D of Ig of bone-marrow-derived lymphocyte strain or hybridoma cell strain secretion because using EBV conversion in vivo still suffer from much asking
Topic, as contained EBV has potential pathogenic；May be with the pathogen such as HIV and hepatitis virus；With hybridoma technology production
The anti-D of Ig is containing Mus source protein, easily cause allergic reaction or with the oncogene of Mus, so by directly injecting Ig in vivo
Anti-D prevents Rh Hemolysis to be just restricted.
Treatment to female tire blood group incompatibility Hemolysis mainly has pregnancy period plasmapheresis, fetal transfusion, termination the most clinically
Gestation and neonatal exchange transfusion are treated.Anti-D titer needs to consider intrauterine transfusion more than 32, and fetal transfusion includes that fetus intraperitoneal is defeated
Blood and the blood transfusion of fetus Ink vessel transfusing, be respectively provided with certain risk, and none be proved effectively；Hyperbilirubinemia of newborn person can use
The Drug therapy such as blue light illumination, immunoglobulin'intravenous, if desired Blood exchanging therapy, Blood exchanging therapy is still molten for treatment neonate
The main method of disorders of blood；During gestation, anti-D titer is more than 64, need to consider replacement therapy of blood plasma, pregnant woman blood plasma displacement be exactly
To filter hemocyte and the blood plasma of method separation anemia of pregnant woman in extracorporeal circulation path, remove blood plasma displacement liquid (the fresh ice with equivalent
Freeze blood plasma or 5% albumin) supplement feedback, each replacement amount 2 000～3 000ml, speed 20～30ml/min, treatment time
2～4h, every 1～3d treatment 1 time；Or take anemia of pregnant woman's whole blood about 400ml every time, remove its blood plasma about 300ml after low-temperature centrifugation, supplement
Amount homotype fresh plasma, the most defeated Autoerythrocyte (RBC).Plasmapheresis proportionally can reduce pathogenic with removed plasma volume
The titre (titer) of antibody, it is thus possible to extend fetus survival and growth and development time in parent, can postpone termination of pregnancy
Time, it is that the anemia of pregnant woman of female tire ABO, Rh or other blood group incompatibility prevents miscarriage the good selection in treatment in early days in clinic, has preferably
Curative effect, also without other untoward reaction.But plasmapheresis can only remove the partial antibody in blood, it is impossible to stop the continuation of antibody
Produce, the female tire blood group incompatibility Hemolysis occurred can not be reversed, need to carry out plasmapheresis incessantly just effectively, the suitableeest
The anemia of pregnant woman of fetus edema, or the homozygote person that spouse is pathogenic antigens was there is, particularly in gestation before 20～22 weeks for once
While removing part pathogenic antibody, the most together eliminate substantial amounts of blood plasma (multiple beneficial composition), although make with displacement liquid
Supplement, but cannot completely supply and be removed blood plasma and various beneficial thereof, and the displacement liquid measure substituted is big, expense is held high
Expensive, supplement allosome blood plasma and easily cause the various side effect such as infectious disease and infusion reaction, which limits generally carrying out of plasmapheresis.
So, only remove pathogenic antibody in the urgent need to one, do not remove blood plasma and multiple beneficial composition thereof, without using blood
Slurry displacement liquid cheap, safely, the treatment new method of female tire blood group incompatibility Hemolysis that has no side effect.
Summary of the invention
In order to solve to attack for a long time the treatment problem of the female tire blood group incompatibility Hemolysis being unable to, present inventors have proposed the present invention.
The invention aims to provide female fetal blood type not conform to blood plasma depurator；Another object is intended to provide the system of depurator
Standby and application process.
The object of the present invention is achieved like this: takes the fresh Rh positive O type erythrocyte of q.s, respectively at 4 DEG C and 37
Clean 4 times with normal saline under conditions of DEG C, to remove erythrocyte surface immune antibody that may be present or natural antibody, so
After with 0.04mol/L Na2HPO4With 0.04mol/L NaH2PO4The PH7.4's that osmotic concentration is 25 and 35mmol/L of preparation
PB lysate replaces Washed Red Blood Cells, finally with normal saline clean until supernatant is colourless, with in erythrocyte of dialysing to human body
Harmful hemoglobin, is centrifuged 10min through 4 DEG C of 2500r/min, and gained cell precipitation is the ghost containing D antigen, through inspection
After surveying cellular morphology and antigenicity, setup of entrances and exits that assembling high-biocompatibility material is made can stop specific particle mesh screen
Cylindrical cleaning device, followed by high speed centrifugation is allowed to be formed the lymphocyte depletion agent of highly dense compression, and depurator top and the bottom are respectively provided with carefully
Born of the same parents' screen cloth, footpath, top is 500 mesh, and footpath, the end is 50 mesh, and liquid outlet arranges the cell strainer of 200 mesh, constitutes and prevents cell debris from entering
Enter the second defence line of circulation, between liquid entrance and mesh screen, relief area is set so that the stablizing, when blood plasma stream of extracorporeal circulation
During purified device, Rh antibody is combined into fixing immune complex by ghost therein, destroyed bib and
Macromole immune complex in combination is cleaned the screen cloth detention of device, and the blood plasma removing morbid substance is filtered from depurator
Feed back internal after going out, remove blood plasma Rh antibody and erythrocytolysis product with this.
Rh antibody is the virulence factor of female tire Rh blood group incompatibility Hemolysis, and Rh (D) positive red blood cell is the natural of Rh antibody
Antigen, the present invention choose the positive O type erythrocyte of Rh (D) make with the dialysis of PB lysate remove be harmful to human body hemoglobin but
Retain the antigenicity of absorption Rh antibody and do not adsorb the ghost of the natural antibody of anti-A and anti-B, be followed by high speed centrifugation system
Become the lymphocyte depletion agent of highly dense compression to be fixed on special depurator with tight intercellular substance, be conducive to compressing cell surface
With filter being fully contacted of blood plasma, thus because adding ghost, the uniform contact face of Rh antibody is increased absorption Rh antibody
Effect, and because being provided with the special screen of cleaner of multiple minimum gateways micropore to bib and macromole antigen
The barrier properties of antibody complex, defines Rh (D) positive ghost absorption Rh antibody and depurator mesh screen mechanical stop is red
The double purified barrier of cell lysates, the erythrocyte flowing through the Rh antibody in the blood plasma of depurator and Hemolysis patient is molten
Hydrolysis products is adsorbed to be removed, and the blood plasma after purification feeds back, it is achieved that manually from internal, Rh antibody is transferred to external treatment, is
One only removes pathogenic antibody and erythrocytolysis thing, do not remove blood plasma and multiple beneficial composition thereof without plasmapheresis liquid
Cheap, safely, the treatment new method of female tire blood group incompatibility Hemolysis that has no side effect.
Detailed description of the invention
Fig. 1 is the application schematic diagram that the female fetal blood type proposed according to the present invention does not conforms to blood plasma depurator.
Fig. 2 is the internal structure schematic diagram of the plasma separator proposed according to the present invention.
Fig. 3 is the internal structure schematic diagram of the depurator 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) being connected with plasma separator (4), plasma separator (4) is through blood plasma pump (6) and circulation line (7) depurator in parallel with 2
(8), depurator (9) be connected, be connected with circulation line (10), venous line (5) the most successively, the other end of venous line (5)
It is connected with vein blood vessel.
In Fig. 2,1 is plasma separator, and 2 is plasma separator inner chamber, and 3 is the micropore on plasma separator inner chamber tube wall, 4
Being the hemocyte that can not pass through micropore (3), 5 is the little molecule blood plasma components that can pass through micropore (3), and 6 is plasma separator exocoel,
7 is blood plasma flow export, and 8 is switchable valve.
In Fig. 3,1 is depurator, the Rh positive ghost that 2 are fixed in depurator, and 3 is the Rh entering depurator
Antibody, 4 is the antigenantibody complex that Rh antibody is formed by Rh positive blood shadow Cell binding, and 5 is RBC fragment.
Below in conjunction with Fig. 1, Fig. 2 and Fig. 3, the female fetal blood type proposing the present invention does not conforms to the embodiment of blood plasma depurator and makees
Detailed description.
One, the preparation of the positive ghost of Rh (D)
1, the acquisition of Rh (D) positive red blood cell
(1) positive " O " the type erythrocyte of fresh concentration Rh (D) is bought from blood station, Zhejiang Province；(2) the most in vitro the giving up of neonate is taken
Abandon positive " O " the type erythrocyte of umbilical cord Rh (D).
2, main agents
30mmol/L PB buffer: solution A is 0.04mol/L Na2HPO4(Na2HPO4.12H2O 14.328g is dissolved in
In 1000ml deionized water, room temperature storage is standby)；Second liquid is 0.04mol/L NaH2PO4(NaH2PO4.2H2O 6.24g is dissolved in
In 1000ml deionized water, room temperature storage is standby).Take respectively solution A 81.0ml mix with second liquid 19.0ml 40mmol/L
PB, then adds deionized water dilution 0.75 times on the basis of 40mmol/L PB buffer and becomes 30mmol/L PB buffering
Liquid, is made into 25 and 35mmol/L in proportion with method.Anti-D standard substance and Rh (D) normal erythrocytes are purchased from the safe biotechnology of Guangzhou connection
Company limited.
3, preparation method
(1) the fresh Rh positive O type erythrocyte of q.s is taken, respectively under conditions of 4 DEG C and 37 DEG C, with 1500r/min
The centrifugal speed of × 5min, cleans erythrocyte 4～5 times with isopyknic normal saline, to remove the possible attachment of 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
The PB lysate of PH7.4, under the centrifugal condition of 4 DEG C and 2500r/min × 10min, alternately and repeatedly Washed Red Blood Cells, pass through
The alternate of osmotic concentration, promotes hemoglobin to give and is removed outside erythrocyte.
(3) finally clean until supernatant is colourless with normal saline, with blood harmful in erythrocyte of dialysing completely
Lactoferrin, precipitation is the ghost containing D antigen.
(4) detection of antigenicity: 1. ghost and anti-D standard substance 37 DEG C are hatched 5min, centrifuging and taking supernatant detects
Anti-D standard substance titer reduces numerical value and determines the antigenicity of ghost.Concrete grammar is: take 10, test tube, numbering 1 respectively
～10, the 1st pipe adds ghost precipitation 1.0ml, and other each pipes add normal saline 0.5ml, then inhale ghost from the 1st pipe
Precipitation 0.5ml joins the 2nd pipe, inhales 0.5ml to the 3rd pipe after mixing, is diluted to the 10th pipe, finally pipe sucking-off with same operation
0.5ml discards, and often pipe adds anti-D standard substance 0.5ml, hatches 5min for 37 DEG C, centrifugal after mix gently, so that complete coagulation, several to occur
Maximum dilution multiple without free cell represents the antigenicity of ghost, and extension rate is the biggest, and antigenicity is the strongest, adsorbs Rh
The ability of antibody is the strongest.Supernatant antibody titer can also be measured with Rh (D) normal erythrocytes and determine the antigen of ghost
Property；2. measuring supernatant antibody titer with Rh (D) normal erythrocytes, anti-D standard substance titer (known) deducts supernatant antibody effect
Valency is the antigenicity (titer) of ghost.Supernatant antibody titer detection method is: take 10 test tubes, respectively numbering 1～
10, each pipe adds normal saline 1ml, takes removal 1ml after supernatant 1ml joins the 1st pipe mixing and goes to the 2nd pipe, with same behaviour
Being diluted to the 10th pipe, finally pipe sucking-off 1ml liquid discards, serum 40 μ l Yu Rh (D) the normal erythrocytes 10 μ l that will have diluted
Mixing, hatches 10 minutes, centrifugal 5 minutes sentence read result, and the coagulation pipe of maximum dilution multiple is antigenicity (titer), and titer should
More than 1: 1500.
(5) content of hemoglobin detection: with hypotonic destruction ghost, with conventional chemical luminescence analysis or fluorescence analysis
Method, the content of hemoglobin in qualitative and detection by quantitative ghost, should be less than the reference value lower bound of human normal plasma.
(6) ghost form: examine under a microscope form and the integrity of ghost, should be in circle shadow, no-reflection.
(7) for cellular morphology in circle shadow, no-reflection, high sensitivity routine hemoglobin qualitative detection is negative, quantitatively
Detection, less than the reference value lower bound of human normal plasma, the ghost of absorption Rh antibody titer more than 1: 1500, is left and taken standby.
Two, the preparation of depurator
1, preparation principle
The present invention is because the free Rh antibody in pregnant woman blood plasma enters fetal blood based on female tire Rh blood group incompatibility Hemolysis
Caused by destruction FE, and Rh antibody can be combined absorption by corresponding native antigen i.e. Rh positive red blood cell, and purifies
The shell of device can be made into the mechanism preparation that the screen cloth only allowing specific little molecule to pass through passes through to stop relatively macromole.
2, material is prepared
Select the high-biocompatibility material close with Human vascular endothelial, it is desirable to good biocompatibility, without complement activation,
NIP reacts, without leukocyte, platelet, blood oxygen pressure, the change of complement C 3 C5a.Require by covalency, be grafted, polymerization etc.
Method improves the uniformity of material surface, hydrophilic, minimizing to blood coagulation and the impact of oxidative stress.Parent is added at depurator inner surface
Hydrogel, as solidified 2 methylacryoyloxyethyl phosphocholines-butyl methacrylate in cellulose acetate membrane, by controlling
Wet-spinning procedure and generate CA/PMB30, CA/PMB80 and CA/PMB30-80, biocompatibility can be improved.By some anticoagulant
Matter is solidificated in carrier or depurator inner surface, and blood coagulation, reduction heparin consumption can be suppressed even to realize no-rod tractor, as by liver
Element is aggregated on polyacrylonitrile-polymine film, can reduce the anaphylaxis of allergic constitution；Heparin covalent is attached to polyethers
Sulfone surface, can keep the mechanical property of polyether sulfone and improve the anticoagulation function of depurator inner surface.On cellulose acetate film altogether
Valency solidification linoleic acid film, maybe will be covalently bound to polyacrylic linoleic acid and be grafted onto polysulfone membrane surface, can have preferably
Histocompatibility and anticoagulant effect.
3, the specification of depurator shell
Make the hydrostatic column that footpath, the end is little, footpath, top is big of 50mm × 60mm, or make square, infundibulate, volume about 200
～300ml, top and the bottom are equipped with cell screen cloth, and footpath, top sieve number is 500 mesh, and footpath, end sieve number is 50 mesh, liquid outlet
Place arranges the cell strainer that mesh number is 200 mesh, constitutes the second defence line preventing cell debris from entering circulation, liquid entrance
And it is provided with relief area, the beneficially stability of system circulation between mesh screen.
4, the filling of cleanser
Take the positive ghost of Rh (D) prepared by the present invention, load depurator, be centrifuged 15～30min with 10000r/min,
Remove supernatant, reinstall ghost, be centrifuged 15～30min with 10000r/min, repeat aforesaid operations, until in depurator
Fill ghost, capping, standby.
Three, the preparation of plasma separator
1, preparation principle: prepare according to the molecular size of hemocyte and blood plasma components.Such as visible component (blood in blood of human body
Cell) size be: normocyte is about 7 microns (μm), is the discoid cell of concave-concave；Leukocyte is divided into 5 kinds, neutral
Granulocyte about 12 μm, eosinophilic granulocyte is more bigger, and basophilic granulocyte is close with neutrophilic granulocyte, small lymphocyte 6-8 μm,
Approximating with erythrocyte, mononuclear cell is maximum, about 15-20 μm.Platelet is disc, and diameter 1～4 microns are to 7～8 microns not
Deng, the platelet mean diameter of people is 2-4 micron, thick 0.5～1.5 micron.
2 prepare material: can be selected for poly-vinegar non-woven fabrics, acetate fiber, absorbent cotton etc., it is desirable to good biocompatibility, hardly
Activating complement, do not cause inflammatory reaction, do not cause the change of leukocyte, platelet, blood oxygen pressure, complement C 3, C5a.Can pass through
Covalency, be grafted, the method such as polymerization improves the structure of material, the regulation microinhomogeneities on surface, hydrophilic, minimizing to blood coagulation and
The impact of oxidative stress thus improve sieving adequacy and biocompatibility, the generation of minimizing complication.
3 specifications: for the profile of separator, can be prepared as cylindricality knot with the material such as acetate fiber or absorbent cotton as filter element
Structure, it is prepared as the shapes such as flat structure as filter element with materials such as poly-vinegar non-woven fabrics；By hemocyte to be separated and blood plasma components
Molecular size determines aperture.Plasma separator stable in properties involved in the present invention, good biocompatibility, height that permeability is high
Molecularly Imprinted Polymer makes hollow fibre type filter, hollow-fiber film a diameter of 270～370 μm, and film thickness is 50 μm, and aperture is
0.2～0.6 μm, fibre length is 13.5～26 μm.This hole is only permitted blood plasma and is filtered, but can stop all of cell component.
Four, the application of the present invention
Extracorporeal circulation branch road need to be collectively constituted with associated components.
1, the component of extracorporeal circulation branch road and purposes
(1) depurator: internal Rh (D) is positive, and ghost is used for removing Rh antibody；The net that depurator shell is imported and exported
Sifter device has the effect filtering the macromolecular complex that RBC fragment, Rh antibody and RBC fragment are formed.
(2) plasma separator: for washed corpuscles and blood plasma.
(3) sound pulse pressure monitoring: arterial blood pressure monitoring mainly in order to the stopping state of dynamic monitoring depurator micropore, is additionally used
With monitoring extracorporeal circulation thrombosis, solidification and the change of pressure.When blood flow deficiency, arterial pressure will reduce；When having blood coagulation, thrombosis
Being formed, particularly during depurator blockage of the micro orifice, arterial pressure will raise；Venous pressure monitoring is used for monitoring the pressure of pipeline blood backflow
Power, when depurator blockage of the micro orifice, blood coagulation, thrombosis, blood flow deficiency and venous return syringe needle come off, venous pressure will
Declining, if the distortion blocking of bloody path return duct or backflow syringe needle occur blocking, venous pressure will raise.
(4) air monitering (Air Detector): be used for monitoring the air bubble of blood pathway, typically use ultrasonic listening
Principle, in order to avoid patient occurs air embolism to arrange.When having monitored air bubble, detecting system can drive dynamic,
Vein bloody path folder carrys out blocking blood flow, prevents the generation of danger.
(5) blood pump (Blood Pump): be used for promoting blood circulation being smoothed out with maintaining treatment, usual blood pump part
Often having rotary test speed function, to monitor the blood circumstance of patient, therefore blood pump runner sets with flute pitch and wants accurately, and
Need often to adjust, according to the situation of bloody path pump line, typically spacing is set as 3.2～3.3mm, can not be the most loose, otherwise can make
Blood flow detection is become to be forbidden；Also can not be too tight, otherwise can cause pipe breakage.
(6) heparin pump (Heparin Pump): heparin pump is equivalent to the micro-injection pump applied clinically, in order to continue to
Injecting heparin in sieving pipeline (patient blood), owing to the blood of patient circulates and air contact in vitro, is susceptible to blood coagulation
Phenomenon, uses heparin pump to be possible to prevent the generation of blood coagulation.
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 a word, on the basis of key constitution system of the present invention, be expected to be further development of automatization, hommization,
Personalization, modularity, automatically monitoring and regulation and control, liquid crystal display, judge the micro computer process such as alarm reason and ring off signal voluntarily
System.
2, the using method of the present invention
(1) install: with sterile working's connecting components, each portion such as including plasma separator, depurator and each circulation line.
(2) aerofluxus: with physiological saline solution topping up separator, depurator and each circulation line, gets rid of separator, depurator
And gas in circulating line, bubble, go through, confirm without use after gas, bubble.
(3) logical liquid: arterial blood line pipe 1 is connected the arteries of HIV sufferers, the most again row of going through
Gas is the most complete, and liquid stream is the most unobstructed, and avoids managing the pollution of interior flow liquid.
(4) anticoagulant: inject anticoagulant (heparin) from heparin pump to liquid stream, be 2500 ∪ or 20～30 ∪/kg for the first time.
(5) start: arterial blood line pipe (1) is connected the arteries of therapist, venous line (5) is connected therapist
Vein blood vessel, then opens blood pump, and blood flow is 100～150ml/min, such as Fig. 1, when arterial blood is through arterial blood line pipe
(1) entering plasma separator (4), the blood plasma separated arrives depurator through circulation line (7) under the effect of blood plasma pump (6)
(8), blood plasma to be full of, about 10 minutes, begin paying out blood plasma, flow out through circulation line (10), synchronize to irrigate blood to depurator (9)
Slurry, when the blood plasma in depurator (8) has nearly flowed, starts again at perfusion blood plasma, and now depurator (9) begins paying out blood plasma,
Two depurators in parallel (8), depurators (9) are alternately.Such as Fig. 2, when blood to be separated enters plasma separator (1)
During inner chamber (2), through the effect of valve (8), little molecule blood plasma and the composition (5) thereof that can pass through micropore (3) enter outside separator
Chamber (6), then flows out through plasma outlet port (7), and the hemocyte (4) that can not pass through micropore (3) flows out through valve (8).Such as Fig. 3,
When Rh antibody (3) enters depurator (1) with blood plasma, Rh positive ghost (2) being cleaned in device is combined into antigen-antibody
Complex (4) and no longer move down, destroyed bib (5) and macromole antigen-antibody in combination are multiple
Mesh screen that compound can not be exported by depurator and by detention, the hemocyte that the blood plasma after purification and plasma separator are separated
Feeding back after converging, so until the plasma circulation amount (usually 9L) being previously set, treatment just ends, if supporting computer
Programme-control, whole therapeutic process is by computer control, and can detect duty at any time, and using can convenient, automatization
And safety.
Five, the checking of practical application of the present invention
In order to verify therapeutic efficiency, the present invention devises the simple method for testing bacterial resistance of depurator effect: take prepared blood shadow
Cell, joins and is incubated after 100 DEG C dissolve in 37 DEG C of 1.0% standby agarose C1-4B, be configured to 95% ghost
The cleanser of concentration；Take 2.5 × 300mm Westergren's blood sedimentation tube 9 of sterilizing, draw the cleanser of 95% ghost concentration extremely
200mm scale, becomes semi-solid after cleanser cooling；Buy blood station, center, Zhejiang Province fresh frozen plasma 200mL, separately buy Rh
Antibody (people's anti-D type serum dry powder standard substance, Guangzhou Lian Tai Bioisystech Co., Ltd), is made into 1: 200,1 with fresh plasma:
300,1: 600 antibody titer, routinely Rh (anti-D) titre detection method (with reference to description), detection confirms above-mentioned institute further
Whether the antibody titer of preparation is consistent, and is referred to as (Rh) antibody (titre) before filter, and before respectively taking 10ml filter, antibody is injected separately into 3
The upper end blank pipe of blood sedimentation tube, after flowing through the adsorbent containing ghost of blood sedimentation tube lower floor, collects effluent, after being referred to as filter
(Rh) antibody, Rh (anti-D) titre detection method confirms titre routinely, more respectively will clean through containing ghost of antibody after filter
Making in agent to leach for the 2nd time, be so repeated 3 times filtration and antibody titer detection, result (table 1) illustrates, Rh antibody filters simple
After cleanser, major part Rh antibody is adsorbed by the cleanser containing ghost accordingly, through the 1st time, the 2nd time, the 3rd filter
Later, the average titer of Rh antibody respectively from filter before 1: 367 reduce to filter after 1: 183,1: 58,1: 29, illustrate along with filtration
The increase of number of times, Rh antibody constantly by ghost adsorption removal, thus can reduce anemia of pregnant woman's (neonate) blood plasma Rh and resist
Body titre and treat the purpose of female tire blood group incompatibility Hemolysis.
Table 1 Rh antibody filters titre testing result (1/x) before and after the cleanser containing ghost
Claims (10)
1. one kind does not conforms to blood plasma depurator for female fetal blood type of medical domain, it is characterised in that prepare two kinds of high and hyposmosis
The PB lysate of concentration, alternately washing Rh positive O type erythrocyte, makes the ghost of hemoglobin of having dialysed, fills in out
The column depurator of screen cloth is set at Kou, prepares the lymphocyte depletion agent of highly dense compression with high speed centrifugation, constitute Rh positive blood shadow thin
The double purified barrier that born of the same parents' absorption and screen of cleaner stop, removes blood plasma Rh antibody and erythrocyte in purging in vitro device
Solute.
Female fetal blood type the most according to claim 1 does not conforms to blood plasma depurator, it is characterised in that described height and hyposmosis concentration
Refer to the PB lysate of the PH7.4 of 25mmol/L and 35mmol/L.
Female fetal blood type the most according to claim 2 does not conforms to blood plasma depurator, it is characterised in that with 81.0ml containing 0.04mol/
L Na2HPO4Solution A and 19.0ml NaH Han 0.04mol/L2PO4Second liquid be made into 40mmol/L after be diluted to deionized water
The PB lysate of the PH7.4 of 25 and 35mmol/L.
Female fetal blood type the most according to claim 1 does not conforms to blood plasma depurator, it is characterised in that described ghost refers to dialysis
Hemoglobin and contain only the characteristic of absorption Rh antibody.
Female fetal blood type the most according to claim 1 does not conforms to blood plasma depurator, it is characterised in that normal saline cleans erythrocyte
Refer to 4 DEG C and 37 DEG C each washings 2 times.
Female fetal blood type the most according to claim 1 does not conforms to blood plasma depurator, it is characterised in that the shell of described depurator holds
Amassing is 200～300ml, and top and the bottom are equipped with cell screen cloth, and footpath, top sieve number is 500 mesh, and footpath, end sieve number is 50 mesh, liquid
Body exit arranges the cell strainer that mesh number is 200 mesh, is provided with relief area between liquid entrance and mesh screen.
Female fetal blood type the most according to claim 1 does not conforms to blood plasma depurator, it is characterised in that separator hollow fibre filter membrane
A diameter of 270～370 μm, film thickness is 50 μm, and aperture is 0.2～0.6 μm, and fibre length is 13.5～26 μm, can filter blood
Starch but all of cell component can not be filtered.
Female fetal blood type the most according to claim 1 does not conforms to blood plasma depurator, it is characterised in that described high speed centrifugation refers to
10000r/min is centrifuged 15～30min.
9. do not conform to blood plasma depurator answering in preparing purging in vitro device according to the arbitrary described female fetal blood type of claim 1-8
With, it is characterised in that described purging in vitro device include one end of arterial blood line pipe (1) through heparin pump (2) and blood pump (3) with
Plasma separator (4) is connected, plasma separator (4) through blood plasma pump (6) and circulation line (7) adsorber (8) in parallel with two,
Adsorber (9) is connected, and is connected with circulation line (10), venous line (5) the most successively.
Application the most according to claim 9, it is characterised in that when enabling the purging in vitro device shown in Fig. 1, tremulous pulse
Blood through arterial blood line pipe (1) enter plasma separator (4), the blood plasma separated under the effect of blood plasma pump (6) through circulation
Pipeline (7) arrives depurator (8), blood plasma to be full of, about 10 minutes, begins paying out blood plasma, flows out through circulation line (10), synchronization
Irrigate blood plasma to depurator (9), when the blood plasma in depurator (8) has nearly flowed, start again at perfusion blood plasma, now depurator
(9) beginning paying out blood plasma, two depurators in parallel (8), depurators (9) are alternately.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109157694A (en) * | 2018-07-01 | 2019-01-08 | 翁炳焕 | A kind of mother's tire blood group incompatibility immunoadsorption therapy instrument |
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| JPS58162295A (en) * | 1982-03-19 | 1983-09-26 | Mitsuru Furusawa | Method for transforming genetic property of animal and vegetable |
| US4752582A (en) * | 1982-12-17 | 1988-06-21 | The United States Of America As Represented By The United States Department Of Energy | Monoclonal antibodies to human glycophorin A and cell lines for the production thereof |
| US20040033232A1 (en) * | 2002-06-13 | 2004-02-19 | Ramberg Elliot R. | Methods and compositions for in vivo clearance of pathogens |
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| CN106267421B (en) | 2019-01-22 |
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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 |