CN106267423A - People's Rh positive red blood cell adsorber - Google Patents

Abstract

The invention relates to an adsorber for human Rh-positive red blood cells in the medical field, which is characterized in that fresh Rh-positive type O red blood cells are washed with physiological saline at 4°C and 37°C to remove immune antibodies or natural antibodies attached to the surface of red blood cells, and after the antigenicity is tested , assemble the cell pellet into a cylindrical adsorber made of highly biocompatible materials to 4/5, then add 3.5% mannitol red blood cell preservation solution to make the red blood cell concentration reach 80%, shake gently to mix, seal, and place for 4 It is stored at ℃ for regular use, and the purifier is equipped with a screen at the outlet to form a line of defense to prevent cell debris from being filtered out. After the plasma is filtered, the Rh antibody is combined by the Rh-positive red blood cells in it to form a fixed complex, and the destroyed red blood cell fragments And the macromolecular complexes combined with it are retained by the screen of the purifier, and the plasma that removes the pathogenic substances is filtered out from the purifier and then reinfused to treat blood group incompatibility hemolytic disease by removing plasma Rh antibodies.

Description

Human Rh positive erythrocyte adsorber

technical field

The invention relates to an adsorber for human Rh-positive red blood cells in the medical field, which is mainly used for removing anti-fetal red blood cell antibodies and red blood cell lysate in plasma of pregnant women with maternal-fetal blood group incompatibility.

Background technique

The fetus inherits half of the genetic components of the father and half of the mother, and the fetal red blood cells can carry antigens from the father, showing that the blood type of the fetus is different from that of the mother. When the fetal red blood cells enter the mother's blood circulation, the mother's immune system is induced to produce antibodies, and the antibodies pass through the placenta and enter the fetal blood circulation system, combining with fetal red blood cells, causing fetal red blood cells to be destroyed, resulting in maternal-fetal blood group incompatibility hemolytic disease in fetuses and newborns .

There are 26 human erythrocyte blood types, including ABO blood type, Rh blood type, and MN, Lew, Kell, and Fya blood types, but Rh blood type and ABO blood type are the most common blood types that can cause maternal-fetal blood group incompatibility hemolytic disease. Rh blood group antigens are determined by 3 pairs of closely linked alleles on chromosome 1, and there are 6 antigens in total, namely C and c, D and d, E and e. Since the D antigen was discovered first and has the strongest antigenicity, in clinical practice, those who are positive for the D antigen are called Rh positive, and those without the D antigen are called Rh negative. The antigenicity of Rh blood group antigens determines the severity of hemolytic disease. D antigen can cause severe hemolytic disease, followed by E antigen, followed by C, c and e antigens. The antigenicity of d antigen is the weakest. At present, there is no anti-d Antibody discovery. The Rh-negative frequency in the Han population in my country is about 0.4%, while the Rh-negative frequency of the ethnic minorities in Northwest China is as high as 10%.

Although the incidence of ABO blood group incompatibility is high, the incidence of fetal hemolysis is very low. Even if hemolysis occurs, the symptoms are mild, kernicterus and edema rarely occur, and no special treatment is required during pregnancy. Although Rh blood group incompatibility is rare, the severity of maternal-fetal blood group incompatibility hemolytic disease caused by it is more serious than that of ABO blood group incompatibility, so the diagnosis and prevention of Rh blood group incompatibility are extremely important. Rh blood group incompatibility is due to the maternal production of a large number of Rh antibodies (IgG anti-D) against fetal red blood cells, which will destroy a large number of fetal red blood cells after entering the fetus, causing severe anemia, hypoxia, heart failure, liver damage, hypoproteinemia, systemic Edema, pleural effusion, ascites and even fetal death. In the neonatal period, Rh blood group incompatibility hemolysis occurs earlier than ABO blood group incompatibility jaundice, the earliest appears within 12 hours after birth, and most of them appear within 24 hours. Because the large amount of bilirubin produced by hemolysis cannot be eliminated from the liver in time, the jaundice of the newborn is aggravated. A large amount of bilirubin penetrates into the brain cells and causes kernicterus. They often die of severe anemia, heart failure, and kernicterus, with a high mortality rate.

Although Rh antibodies can cause severe hemolytic disease by destroying fetal red blood cells after entering the fetus, some pregnant women can also prevent the invading Rh(D) positive red blood cells from sensitizing the body by pre-injecting Rh antibodies, thereby preventing Rh hemolysis the occurrence of disease. Since most countries have banned the use of volunteers for D antigen immunization, the source of polyclonal Ig anti-D is very limited, and the use of Epstein-Barr virus to transform human B lymphocytes to secrete Ig anti-D or Epstein-Barr virus transformation combined with cell fusion to prepare Ig Anti-D, because the use of Ig anti-D secreted by EBV-transformed B lymphocyte lines or hybridoma cell lines in vivo still has many problems, such as the potential pathogenicity of EBV contained; it may carry HIV and hepatitis viruses and other pathogens; the Ig anti-D produced by hybridoma technology contains mouse protein, which is easy to cause allergic reactions, or contains mouse oncogenes, so the prevention of Rh hemolytic disease by direct injection of Ig anti-D in the body is limited .

At present, the clinical treatment of maternal-fetal blood group incompatibility hemolytic disease mainly includes plasma exchange during pregnancy, fetal blood transfusion, termination of pregnancy and neonatal exchange transfusion. Intrauterine blood transfusion should be considered if the anti-D titer is above 32. Fetal blood transfusion includes fetal intraperitoneal blood transfusion and fetal intravascular blood transfusion, both of which have certain risks, and none of them has been proven effective; neonatal hyperbilirubinemia can be used Blu-ray irradiation, intravenous immunoglobulin and other drug treatments, exchange transfusion if necessary, exchange transfusion is still the main method for the treatment of hemolytic disease of the newborn; during pregnancy, if the anti-D titer is above 64, plasma exchange therapy should be considered, and plasma exchange in pregnant women In the extracorporeal circulation channel, the blood cells and plasma of pregnant women are separated by filtration, the plasma is removed, and an equal amount of replacement fluid (fresh frozen plasma or 5% albumin) is added and reinfused. The speed is 20-30ml/min, the treatment time is 2-4 hours, and the treatment is performed once every 1-3 days; or about 400ml of whole blood from pregnant women is taken each time, and about 300ml of plasma is removed after low-temperature centrifugation. Red blood cells (RBCs). Plasma exchange can reduce the titer (titer) of pathogenic antibodies in proportion to the amount of plasma removed, thereby prolonging the survival and growth and development time of the fetus in the mother, and delaying the time of termination of pregnancy. It is the mother-fetal ABO, Rh It is a good choice for early clinical miscarriage treatment for pregnant women with incompatibility or other blood types. It has good curative effect and no other adverse reactions. However, plasma exchange can only remove part of the antibodies in the blood, and cannot stop the continuous production of antibodies, nor can it reverse the existing maternal-fetal blood group incompatibility hemolytic disease. It needs continuous plasma exchange to be effective. Pregnant women who had fetal hydrops two weeks ago, or whose spouses are homozygous for the pathogenic antigen, especially when removing part of the pathogenic antibody, also remove a large amount of plasma (multiple beneficial components), although the replacement fluid Supplementation has been made, but the removed plasma and its various beneficial components cannot be completely replenished, and the replacement fluid for the replacement is large and expensive. Supplementing allogeneic plasma is prone to various side effects such as infectious diseases and infusion reactions, which limits plasma exchange. widespread development.

Therefore, there is an urgent need for a new method for the treatment of maternal-fetal blood incompatibility hemolytic disease that only removes pathogenic antibodies, does not remove plasma and its various beneficial components, and does not need to use plasma replacement fluid, which is cheap, safe, and has no side effects.

Contents of the invention

In order to solve the problem of the treatment of maternal-fetal blood group incompatibility hemolytic disease, which cannot be overcome for a long time, the inventors proposed the present invention.

The object of the present invention is to provide an adsorber for human Rh positive red blood cells; another object is to provide a preparation and application method of the adsorber.

The object of the present invention is achieved in this way: according to the collection requirements of component blood transfusion by the blood collection and supply institution, fresh Rh-positive "O" type concentrated red blood cells are prepared, and they are washed 4 times with normal saline at 4°C and 37°C respectively to remove Immunity antibodies or natural antibodies that may exist on the surface of red blood cells, after testing the antigenicity, take the cell pellet and assemble a cylindrical adsorber made of high biocompatibility materials, to 4/5, add 3.5% mannitol red blood cell preservation solution, Make the concentration of red blood cells reach 80%, shake gently to mix, seal, store at 4°C for later use, install cell screens on the upper and lower parts of the adsorber, the top diameter is 500 mesh, the bottom diameter is 50 mesh, and the liquid outlet is set with 200 mesh cells The filter screen constitutes the second line of defense to prevent cell debris from entering the circulation. A buffer zone is set between the liquid inlet and outlet and the screen to stabilize the extracorporeal circulation. When the plasma flows through the adsorber, the Rh antibody is bound by the Rh-positive red blood cells in it. The damaged red blood cell fragments and the macromolecular immune complexes combined with them are retained by the screen of the adsorber, and the plasma that removes the pathogenic substances is filtered out from the adsorber and then returned to the body. This clears plasma Rh antibodies and erythrocyte lysates.

Rh antibody is the pathogenic factor of maternal-fetal Rh blood incompatibility hemolytic disease, and Rh (D) positive red blood cells are the natural antigen of Rh antibody, which can be combined with Rh antibody to remove it by adsorption. Wash with normal saline at room temperature to remove natural and immune blood group antibodies on the surface of red blood cells, but retain the antigenicity of adsorbed Rh antibodies without adsorbing natural antibodies against-A and anti-B, followed by 3.5% mannitol red blood cell preservation solution The medium is equipped with an adsorber, which is easy to store regularly, Rh(D) positive red blood cells are easy to obtain, the preparation method is simple, and the special adsorber screen with multiple extremely small inlet and outlet micropores can compound red blood cell fragments and macromolecular antigens and antibodies The barrier properties of substances form a double purification barrier for Rh positive red blood cells to adsorb Rh antibodies and the adsorber mesh mechanically blocks red blood cell lysates, so that the Rh antibodies in the plasma flowing through the adsorber and the red blood cell lysates of patients with hemolytic diseases are adsorbed and removed. , the purified plasma is reinfused, so as to realize the treatment of removing Rh antibodies from the body. It is a cheap blood replacement solution that only removes pathogenic antibodies and erythrocyte lysates, does not remove plasma and its various beneficial components, and does not require plasma replacement fluid. A new method for the treatment of maternal-fetal blood group incompatibility hemolytic disease, which is safe and has no side effects.

detailed description

Fig. 1 is a schematic diagram of the application of the human Rh positive red blood cell adsorber proposed according to the present invention.

Fig. 2 is a schematic diagram of the internal structure of the plasma separator proposed according to the present invention.

Figure 3 is a schematic diagram of the internal structure of the immunoadsorber proposed according to the present invention

In Fig. 1, one end of the arterial blood line (1) is connected with the arterial blood vessel, and the other end is connected with the plasma separator (4) through the heparin pump (2) and the blood pump (3), and the plasma separator (4) is connected with the plasma separator (4) through the plasma pump. (6) and the circulation pipeline (7) are connected with two parallel adsorbers (8) and adsorbers (9), and then connected with the circulation pipeline (10) and the vein pipeline (5) successively, and the vein pipeline ( 5) The other end is connected with the vein blood vessel.

In Fig. 2, 1 is the plasma separator, 2 is the inner cavity of the plasma separator, 3 is the micropore on the wall of the plasma separator inner cavity, 4 is the blood cell that cannot pass through the micropore (3), and 5 is the micropore that can pass through the micropore (3) The small molecule plasma component, 6 is the outer cavity of the plasma separator, 7 is the plasma outflow port, and 8 is a switchable valve.

In Figure 3, 1 is the adsorber, 2 is the Rh-positive red blood cells located in the adsorber, 3 is the Rh antibody entering the adsorber, 4 is the antigen-antibody immune complex formed by the Rh antibody being bound by Rh-positive red blood cells, and 5 is RBC debris.

The embodiment of the human Rh positive erythrocyte adsorber proposed by the present invention will be described in detail below with reference to FIG. 1 , FIG. 2 and FIG. 3 .

1. Preparation of Rh(D) positive red blood cells

1. Acquisition of Rh(D) positive red blood cells

(1) Purchasing fresh concentrated Rh(D) positive "O" red blood cells from blood collection and supply institutions in Zhejiang Province; (2) Extracting Rh(D) positive "O" red blood cells from the discarded umbilical cord blood of newborns.

2. Main reagents and their functions

The formula of 3.5% mannitol red blood cell preservation solution is: take 17.5g mannitol, 1.5g trisodium citrate, 0.2g citric acid, 7.93g glucose, 0.94g sodium dihydrogen phosphate, 0.14g adenine, sodium chloride 4.97g, dissolved in 500ml of distilled water. Among them, mannitol dilutes the viscosity of concentrated red blood cells, increases the stability of cell membranes, and prevents hemolysis. Sodium citrate combines with Ca2+ in blood or plasma to form soluble calcium citrate that is difficult to dissociate, reducing Ca2+ in blood, thereby inhibiting the coagulation process, producing anticoagulant effect, and having a protective effect on red blood cells to prevent hemolysis occur. Citric acid prevents the coking of glucose in the red blood cell supplement, and forms a buffer pair with sodium citrate to adjust and stabilize the pH of the solution. Glucose is the main energy source for red blood cell metabolism. Under normal circumstances, 90% of glucose is anaerobic glycolysis, and 10% is generated through the pentose phosphate pathway to generate ATP, which provides energy for red blood cells to maintain their lifespan. Adenine increases blood ATP activity levels during storage at 4°C. Red blood cells have a specific need for adenine, which can convert adenine into adenosine phosphate (AMP), and further phosphorylate to generate ATP, which provides a source of high-energy compounds for red blood cell metabolism and greatly prolongs the storage time of blood at 4°C. Sodium chloride adjusts the osmotic pressure of the solution to be an isotonic solution, providing an appropriate amount of sodium ions for red blood cells. Sodium dihydrogen phosphate prevents red blood cell aggregation, provides phosphate for red blood cell energy metabolism, and slows down the decline rate of 2,3-DPG.

Anti-D standard and Rh(D) standard red blood cells were purchased from Guangzhou Liantai Biotechnology Co., Ltd.

3. Preparation method

(1) Fresh Rh-positive "O"-type concentrated red blood cells are prepared by the blood collection and supply institution according to the collection requirements of component blood transfusion, and a sufficient amount of fresh Rh-positive type O red blood cells is taken, and the temperature is 4°C and 37°C, respectively, at a rate of 1500r/ Min × 5min centrifugation speed, wash the red blood cells with equal volume of normal saline 4-5 times to remove the ABO blood type and Rh blood type and other immune antibodies or natural antibodies that may be attached to the surface of the red blood cells.

(2) Antigenicity detection: ①Incubate erythrocytes with anti-D standard substance at 37°C for 5 minutes, centrifuge and take the supernatant to detect the decrease in titer of anti-D standard substance to determine the antigenicity of erythrocytes. The specific method is: take 10 test tubes, numbered 1 to 10 respectively, add 1.0ml of red blood cell pellet to the first tube, add 0.5ml of normal saline to the other tubes, then suck 0.5ml of red blood cell pellet from the first tube and add it to the second tube, mix After uniformity, suck 0.5ml to the third tube, dilute to the tenth tube by the same operation, suck out 0.5ml from the last tube and discard, add 0.5ml anti-D standard to each tube, incubate at 37°C for 5min, centrifuge and mix gently. The antigenicity of erythrocytes is represented by the maximum dilution factor at which complete agglutination occurs and there are almost no free cells. The greater the dilution factor, the stronger the antigenicity and the stronger the ability to adsorb Rh antibodies. The antigenicity of red blood cells can also be determined by measuring the antibody titer of the supernatant with Rh (D) standard red blood cells; Subtracting the supernatant antibody titer is the antigenicity (titer) of red blood cells. The method for detecting the antibody titer of the supernatant is: take 10 test tubes, numbered 1-10 respectively, add 1ml of normal saline to each tube, take 1ml of the supernatant and add it to the first tube, mix well, remove 1ml and transfer it to the second tube, Dilute to the 10th tube by the same operation, suck out 1ml of the liquid at the end and discard it, mix 40μl of the diluted serum with 10μl of Rh(D) standard red blood cells, incubate for 10 minutes, and centrifuge for 5 minutes to interpret the results. The agglutination tube with the maximum dilution factor is Antigenicity (titer), the titer should be above 1:1500.

2. Preparation of the adsorber

1. Preparation principle

The present invention is based on the fact that maternal-fetal Rh blood group incompatibility hemolytic disease is caused by the free Rh antibody in the pregnant woman's plasma entering the fetal blood to destroy fetal red blood cells, and the Rh antibody can be bound and adsorbed by the corresponding natural antigen, that is, Rh-positive red blood cells, and the shell of the adsorber can be made It is prepared by a mechanism that only allows specific small molecules to pass through, while blocking the passage of larger molecules.

2. Preparation materials

Highly biocompatible materials close to human vascular endothelium are selected, which require good biocompatibility, no complement activation, no inflammatory response, no changes in leukocytes, platelets, blood oxygen partial pressure, and complement C3aC5a. It is required to improve the uniformity and hydrophilicity of the material surface through covalent, grafting, polymerization and other methods, and reduce the impact on blood coagulation and oxidative stress. Add hydrophilic gel on the inner surface of the adsorber, such as curing 2 methacryloyloxyethyl phosphorylcholine-butyl methacrylate on cellulose acetate membrane, and generate CA/PMB30, CA/PMB80 by controlling the wet spinning process And CA/PMB30-80, can improve biocompatibility. Immobilizing some anticoagulant substances on the inner surface of the carrier or adsorber can inhibit blood coagulation, reduce the amount of heparin and even realize heparin-free. For example, polymerizing heparin on polyacrylonitrile-polyethyleneimine membrane can reduce the risk of allergic Anaphylaxis; covalent binding of heparin to the surface of polyethersulfone can maintain the mechanical properties of polyethersulfone and improve the anticoagulant performance of the inner surface of the adsorber. Covalently curing linoleic acid membranes on cellulose acetate membranes, or grafting linoleic acid covalently bound to polyacrylic acid onto the surface of polysulfone membranes, can have better histocompatibility and anticoagulant effects.

3. Specifications of the adsorber shell

Made into a cylindrical container with a small bottom diameter and a large top diameter of 50mm×60mm, or made into a square or funnel shape, with a volume of about 200-300ml, and cell screens are installed on the upper and lower parts, and the mesh number of the top diameter screen is 500 mesh , the bottom diameter screen mesh is 50 mesh, and the cell filter with a mesh number of 200 mesh is set at the liquid outlet, which constitutes the second line of defense to prevent cell debris from entering the circulation. There is a buffer zone between the liquid inlet and outlet and the mesh screen. Conducive to the stability of the system cycle.

4. Filling of adsorbent

Take the Rh(D) positive erythrocytes prepared by the present invention, put them into an adsorber, and add 3.5% mannitol erythrocyte preservation solution to make the erythrocyte concentration reach 80%, shake gently to mix, seal, and store at 4°C spare.

3. Preparation of Plasma Separator

1. Preparation principle: It is prepared according to the molecular size of blood cells and plasma components. For example, the size of formed components (blood cells) in human blood is: normal red blood cells are about 7 microns (μm), which are biconcave disc-shaped cells; white blood cells are divided into 5 types, neutrophils are about 12 μm, and eosinophils are slightly smaller. Larger, basophils are close to neutrophils, small lymphocytes are 6-8 μm, similar to red blood cells, and monocytes are the largest, about 15-20 μm. Platelets are disc-shaped, with a diameter ranging from 1 to 4 microns to 7 to 8 microns. The average diameter of human platelets is 2-4 microns and a thickness of 0.5 to 1.5 microns.

2 Preparation materials: Polyester non-woven fabrics, cellulose acetate, absorbent cotton, etc. can be used, which require good biocompatibility, almost no activation of complement, no inflammatory response, no leukocytes, platelets, blood oxygen partial pressure, complement C3a, C5a change. The structure of the material can be improved by means of covalent, grafting, polymerization, etc., and the microscopic heterogeneity and hydrophilicity of the surface can be adjusted to reduce the impact on blood coagulation and oxidative stress, thereby improving the adequacy of screening and biocompatibility. Reduce the occurrence of complications.

3 Specifications: As far as the shape of the separator is concerned, it can be prepared into a columnar structure with acetate fiber or absorbent cotton as the filter element, and can be prepared into a flat structure with polyester non-woven fabric as the filter element; according to the blood cells and plasma to be separated The molecular size of the constituents determines the pore size. The plasma separator involved in the present invention is made of a high molecular polymer with stable properties, good biocompatibility and high permeability to make a hollow fiber filter. The diameter of the hollow fiber membrane is 270-370 μm, the membrane thickness is 50 μm, and the pore size is 0.2 μm. ～0.6 μm, fiber length is 13.5～26 μm. The pores permit only plasma filtration but block all cellular components.

Four, application of the present invention

It needs to form an extracorporeal circulation branch together with relevant components.

1. Components and uses of extracorporeal circulation branch

(1) Adsorber: the Rh (D) positive red blood cells inside are used to remove Rh antibodies; the mesh screen at the inlet and outlet of the adsorber shell has the function of filtering out RBC fragments and macromolecular complexes formed by Rh antibodies and RBC fragments.

(2) Plasma separator: used to separate blood cells and plasma.

(3) Arterial and venous pressure monitoring: Arterial pressure monitoring is mainly used to dynamically monitor the blockage of the micropores of the adsorber, and to monitor changes in extracorporeal circulation thrombus, coagulation and pressure. When the blood flow is insufficient, the arterial pressure will decrease; when there is coagulation and thrombus formation, especially when the micropores of the adsorber are blocked, the arterial pressure will increase; When the pores of the device are blocked, coagulation, thrombosis, insufficient blood flow, and the venous return needle falls off, the venous pressure will drop. If the blood return tube is twisted and blocked or the return needle is blocked, the venous pressure will increase.

(4) Air Detector: It is used to monitor the air bubbles in the blood flow path, generally using the principle of ultrasonic detection, and it is set up to avoid air embolism in patients. When air bubbles are detected, the detection system will drive arterial and venous blood clamps to block blood flow and prevent danger.

(5) Blood Pump: It is used to promote blood circulation to maintain the smooth progress of the treatment. Usually, the blood pump part often has the function of speed detection to monitor the blood flow of the patient. Therefore, the distance between the blood pump runner and the groove is set It must be accurate and needs to be adjusted frequently. According to the condition of the blood pump tube, the spacing is generally set at 3.2-3.3mm. It should not be too loose, otherwise it will cause inaccurate blood flow detection; rupture.

(6) Heparin Pump: The heparin pump is equivalent to a clinically used micro-injection pump, which is used to continuously inject heparin into the screening pipeline (patient's blood). Coagulation phenomenon, the use of heparin pump can prevent the occurrence of coagulation.

In addition, it also includes temperature control system, liquid distribution system, degassing system, conductivity monitoring system, ultrafiltration monitoring and blood leakage monitoring and other parts. In a word, on the basis of the key constituent system of the present invention, it is expected to be further developed into a microcomputer processing system such as automation, humanization, individualization, modularization, automatic monitoring and control, liquid crystal display, self-judging the cause of the alarm and releasing the signal.

2, the use method of the present invention

(1) Installation: connect various parts by aseptic operation, including plasma separator, adsorber and various circulation lines.

(2) Exhaust: Fill the separator, adsorber and each circulation pipeline with sterile saline, remove the gas and air bubbles in the separator, adsorber and their circulation pipeline, check carefully, and use it after confirming that there is no gas or bubble .

(3) Fluid connection: connect the arterial blood tube 1 to the arterial blood vessel of the AIDS patient, and carefully check again during the operation whether the exhaust is complete, whether the liquid flow is unobstructed, and to avoid contamination of the fluid in the tube.

(4) Anticoagulation: Inject anticoagulant (heparin) from the heparin pump into the liquid flow, the initial rate is 2500∪ or 20～30∪/kg.

(5) Start: connect the arterial blood line (1) to the arterial blood vessel of the patient, connect the venous line (5) to the venous blood vessel of the patient, and then turn on the blood pump. The blood flow rate is 100-150ml/min. Figure 1, when the arterial blood enters the plasma separator (4) through the arterial blood line (1), the separated plasma reaches the adsorber (8) through the circulation line (7) under the action of the plasma pump (6), and waits to be filled Plasma, about 10 minutes, start to release plasma, flow out through the circulation line (10), and simultaneously perfuse plasma into the adsorber (9), when the plasma in the adsorber (8) is nearly exhausted, start perfusing plasma again, at this time The adsorber (9) starts to release plasma, and the two parallel adsorbers (8) and adsorbers (9) are alternately performed. As shown in Figure 2, when the blood to be separated enters the inner cavity (2) of the plasma separator (1), through the action of the valve (8), the small molecule plasma and its components (5) can enter through the micropore (3) The outer cavity (6) of the separator then flows out through the plasma outlet (7), while the blood cells (4) that cannot pass through the micropore (3) flow out through the valve (8). As shown in Figure 3, when the Rh antibody (3) enters the adsorber (1) with the plasma, the Rh-positive red blood cells (2) in the adsorber combine to form an antigen-antibody complex (4) and no longer move down, and the destroyed Red blood cell fragments (5) and the macromolecular antigen-antibody complexes combined with them cannot be blocked by the mesh screen at the outlet of the adsorber, and the purified plasma is reinfused with the blood cells separated from the plasma separator, and so on until The pre-set plasma circulation volume (usually 9L) is the end of the treatment. If it is controlled by a computer program, the entire treatment process is controlled by the computer, and the working status can be checked at any time. The use will be more convenient, automatic and safe.

Five, the verification of the application effect of the present invention

In order to verify the efficacy of the adsorber in removing Rh antibodies, the present invention has designed a simple test method: take 9 sterilized 2.5×300mm Widmanner tubes, absorb Rh (D) positive erythrocytes to precipitate to the 250mm scale, and then absorb an appropriate amount of The 1.0% agarose C1-4B that is kept at 42°C for standby after melting at 100°C, the agarose becomes semi-solid after cooling, can fix red blood cells in the tube, and make a simple cylindrical adsorber. In addition, 200 mL of fresh frozen plasma and Rh antibody (human anti-type D serum dry powder standard product, Guangzhou Liantai Biotechnology Co., Ltd.) were purchased from the Central Blood Bank of Zhejiang Province, and fresh plasma was used to prepare 1:200, 1:300, and 1:600 Antibody titer, according to the conventional Rh (anti-D) titer detection method (refer to the instruction manual), further testing to confirm whether the above-mentioned prepared antibody titer is consistent, called pre-filtered (Rh) antibody (titer), and then each took 10ml The pre-filtered antibodies were injected into the upper empty tubes of the three erythrocyte sedimentation tubes, and after the red blood cells in the erythrocyte sedimentation tubes flowed through, the effluent was collected, which was called the filtered (Rh) antibody, and the titer was confirmed according to the conventional Rh (anti-D) titer detection method , and then filter the filtered antibody for the second time through the blood sedimentation tube containing Rh (D) positive red blood cells, and repeat the filtering and antibody titer detection for 3 times. The results (Table 1) show that the Rh antibody is easy to filter After the adsorber, most of the Rh antibodies have been adsorbed by the corresponding Rh(D) positive red blood cells. After the first, second, and third filtration, the average titers of Rh antibodies increased from 1:367 to 1:367 before filtration. Reduced to 1:183, 1:58, and 1:29 after filtration, indicating that with the increase in the number of filtrations, Rh antibodies will be continuously removed by Rh (D) positive red blood cells, thereby achieving a reduction in maternal (newborn) plasma Rh antibody titers for the purpose of treating maternal-fetal blood group incompatibility hemolytic disease.

Table 1 The titer detection results before and after Rh antibody filtration of the adsorber containing Rh(D) positive red blood cells (1/x)

Claims (10)

1. A human Rh-positive red blood cell adsorber used in the medical field, characterized in that Rh-positive "O" type red blood cells are washed with 4°C and 37°C normal saline respectively to remove immunity and natural antibodies, and 3.5% manna Alcoholic erythrocyte preservation solution is used to prepare cell suspension with 80% concentration, and the columnar adsorber with screen mesh at the perfusion outlet is used to remove plasma Rh antibody and erythrocyte lysate in the in vitro adsorption device. 2. The human Rh positive erythrocyte adsorber according to claim 1, wherein the Rh antibodies refer to Ig anti-D, Ig anti-E, Ig anti-e, Ig anti-C, and Ig anti-c. 3 . The human Rh positive red blood cell adsorber according to claim 1 , wherein the Rh positive "O" type red blood cells can only adsorb Rh antibodies. 4. The human Rh positive erythrocyte adsorber according to claim 1, characterized in that, the volume of the shell of the adsorber is 200-300ml, the upper and lower parts are provided with cell screens, and the mesh number of the top diameter screen is 500 mesh , the bottom diameter screen mesh is 50 mesh, the liquid outlet is provided with a cell strainer with a mesh number of 200 mesh, and a buffer zone is provided between the liquid inlet and outlet and the mesh screen. 5 . The human Rh positive erythrocyte adsorber according to claim 1 , wherein the extracorporeal adsorption device comprises a plasma separator. 6 . The human Rh positive erythrocyte adsorber according to claim 5 , wherein the plasma separator is a hollow fiber filter, and the inner cavity communicates with the outer cavity through micropores on the tube wall. 7. The human Rh positive erythrocyte adsorber according to claim 6, characterized in that the hollow fiber filter membrane of the separator has a diameter of 270-370 μm, a membrane thickness of 50 μm, a pore diameter of 0.2-0.6 μm, and a fiber length of 13.5 μm. ~26 μm, can filter plasma but not all cellular components. 8. The human Rh-positive erythrocyte adsorber according to claim 1, wherein the erythrocyte preservation solution of 3.5% mannitol is 17.5g of mannitol, 1.5g of trisodium citrate, 0.2g of citric acid, Glucose 7.93g, sodium dihydrogen phosphate 0.94g, adenine 0.14g, sodium chloride 4.97g, dissolved in 500ml of distilled water. 9. The application of the human Rh positive erythrocyte adsorber according to any one of claims 1-7 in the preparation of an in vitro adsorption device, characterized in that, the in vitro adsorption device comprises an end of the arterial blood line (1) via a heparin pump ( 2) The blood pump (3) is connected to the plasma separator (4), and the plasma separator (4) is connected to two parallel adsorbers (8), adsorber ( 9) to be connected, and then connected to the circulation line (10) and the venous line (5) in sequence. 10. The application according to claim 9, characterized in that, when the in vitro adsorption device shown in Figure 1 is enabled, the arterial blood enters the plasma separator (4) through the arterial blood line (1), and the separated plasma is in the plasma Under the action of the pump (6), it reaches the adsorber (8) through the circulation pipeline (7). After being filled with plasma, it starts to release the plasma for about 10 minutes, flows out through the circulation pipeline (10), and simultaneously pours into the adsorber (9) Plasma, when the plasma in the adsorber (8) is about to flow out, start to perfuse the plasma again, at this moment, the adsorber (9) begins to emit plasma, and two parallel adsorbers (8), adsorbers (9) alternately carry out.