Method of purulent inflammatory complication prevention for orthopedic trauma cases, using autologic lymphocyte culture

FIELD: medicine.

SUBSTANCE: invention concerns medicine, particularly traumatology and orthopedics, and can be applied as preventive measure against purulent inflammatory complications for orthopedic trauma cases. It involves whole blood take from any central or peripheral vein of patient on the day before surgical intervention in amount of 10 ml by syringe flushed by heparin in advance. Obtained whole blood is settled for 100-140 minutes, lymphocyte fraction is separated in amount of 0.5-1.5 ml with further adding 5 ml of RPMI-1640 cultural medium and 5 ml of the patient's own serum to the fraction. Obtained cell suspension is poured by 2 ml into sterile vessels, and 5 ml of RPMI-1640 cultural medium and 0.01 ml of 10% phytohemoagglutinin solution in RPMI-1640 cultural medium is added to each vessel. Vessels with lymphocyte culture are placed in thermostat and kept at 37°C for 72 hours. Then autologic lymphocyte culture obtained in each vessel is centrifuged for 3-5 times for 10 minutes at 800-1200 rpm rotation rate. After each centrifugation, supernatant fluid is discharged, and 5 ml of saline solution is added to each vessel for repeated centrifugation. When centrifugation is over, cell bulk of autologic lymphocytes is gathered from all vessels by a syringe with added 10 ml of saline solution, and obtained bulk is further injected intravenously to any central or peripheral vein for 30-70 seconds. Cell bulk obtained by the method is injected before surgical intervention, or during it, or over 2-3 days after surgical intervention. Cell bulk of autologic lymphocytes can also be performed again over 7-14 days after initial injection.

EFFECT: enhanced immune organism reactivity in post-operation period due to increased lymphocyte number in blood.

2 cl, 3 ex

 

The invention relates to medicine, namely to traumatology, orthopedics and rehabilitation, to methods of prevention of inflammatory complications in trauma and orthopaedic patients using culture autologous lymphocytes, and can be used for the prevention of inflammatory complications in patients in surgical, trauma and other hospitals.

There is a method of treatment of purulent-inflammatory complications in trauma and orthopaedic patients using culture autologous lymphocytes, including the blood of the patient, the cultivation of lymphocytes and intravenous their patient (see Gagasan and other Remodeling protective properties of the body with extensive operational interventions in traumatology and orthopedics. Medical Bulletin of Erebuni, the national Academy of Sciences of Armenia, Yerevan, 2006, №3 (27), p.140-141).

However, the known method when its use has the following disadvantages:

- does not provide reliable treatment and prevention of inflammatory complications in trauma and orthopedic patients after suffering a high surgical aggression

- does not provide a sufficiently favorable for traumatic disease,

- does not provide necessary and sufficient increase in the quantities of the lymphocytes in the blood of a patient after suffering a high surgical aggression

- not enough provides increased cellular immunity

- not enough provides the patient the necessary increase of immunoglobulins.

The objective of the invention is to provide a method of prophylaxis of purulent-inflammatory complications in trauma and orthopaedic patients using culture autologous lymphocytes.

The technical result is the ability to provide reliable prevention of inflammatory complications in trauma and orthopedic patients after undergoing surgery, providing necessary and sufficient increase in the number of lymphocytes in the blood of a patient after undergoing surgery, as well as providing enough of a favorable course of traumatic disease of the patient after suffering a high surgical aggression. In addition, the technical result is to provide the patient the necessary increase of immunoglobulins.

The technical result is achieved by the fact that in the proposed method of prevention of inflammatory complications in trauma and orthopaedic patients using culture autologous lymphocytes intake of whole blood of a patient before surgical intervention of any Central or peripheral vein in pre-Amity heparin syringe of 10 ml and defend it within 100-140 minutes select lymphocytic fraction in the amount of 0.5-1.5 ml and injected into 5 ml of culture medium RPMI-1640 and 5 ml of private patient serum is poured obtained cell suspension in sterile containers 2 ml and injected in every capacity 5 ml of culture medium RPMI-1640 and 0.01 ml of 10% solution of phytohemagglutinin in culture medium RPMI-1640, capacity, culture of lymphocytes placed in an incubator and incubated at 37°C for 72 hours, then obtained in each tank cell culture autologous lymphocytes centrifuged 3-5 times at a speed of rotation 1000 rpm for 10 minutes, after each centrifugation the supernatant is poured the liquid in each tank add 5 ml of saline to repeat the centrifugation of the cell culture with autologous lymphocytes, then after centrifugation removed from all tanks in the syringe-cell mass autologous lymphocytes, in a syringe with typed cell mass autologous lymphocytes add 10 ml of saline and administered intravenously to the patient in any Central or peripheral vein for 30-70 seconds, with intravenous patient cell mass autologous lymphocytes in the physiological solution is performed before performing surgery, or intraoperation is, or 2-3 days after surgery. While intravenous patient cell culture of autologous cells in physiological solution additionally perform repeatedly after 7-14 days after initial injection.

The method is as follows. Before surgical intervention perform the sampling of whole blood of the patient from any Central or peripheral vein in a pre-rinsed with heparin syringe of 10 ml of Collected whole blood of the patient advocate within 100-140 minutes. Select lymphocytic fraction in the amount of 0.5-1.5 ml and injected into 5 ml of culture medium RPMI-1640 and 5 ml of private patient serum. The obtained cell suspension is poured into sterile containers 2 ml and injected in every capacity 5 ml of culture medium RPMI-1640 and 0.01 ml of 10% solution of phytohemagglutinin in culture medium RPMI-1640. Capacity with the culture of lymphocytes placed in an incubator and incubated at 37°C for 72 hours. Received in each tank cell culture autologous lymphocytes centrifuged 3-5 times at a speed of 800-1200 rpm for 10 minutes each centrifugation. After each centrifugation the supernatant is poured the liquid in each tank add 5 ml of physiological solution for having overenia centrifugation of the cell culture with autologous lymphocytes. After centrifugation removed from all tanks in the syringe-cell mass autologous lymphocytes, in a syringe with typed cell mass autologous lymphocytes add 10 ml of saline and administered intravenously to the patient in any Central or peripheral vein for 30-70 seconds. While intravenous patient cell mass autologous lymphocytes in the physiological solution is performed before performing surgery, or intraoperatively, or 2-3 days after complete surgical intervention. Intravenous patient cell culture of autologous cells in physiological solution additionally perform repeatedly after 7-14 days after initial injection.

Among the essential features characterizing the proposed method of prophylaxis of purulent-inflammatory complications in trauma and orthopaedic patients using culture autologous lymphocytes, distinctive are:

- exercise intake of whole blood of the patient before surgical intervention of any Central or peripheral vein in a pre-rinsed with heparin syringe of 10 ml, and advocating for 100-140 minutes

- selection lymphocytic fraction of the patient in the amount of 0.5-1.5 ml and the introduction is not the 5 ml culture medium RPMI-1640, and 5 ml of private patient serum

- pouring the obtained cell suspension in sterile containers 2 ml and the introduction of in every capacity 5 ml of culture medium RPMI-1640 and 0.01 ml of 10% solution of phytohemagglutinin in culture medium RPMI-1640,

- placement of containers with the culture of lymphocytes in thermostat and keeping at a temperature of 37°C for 72 hours,

- performance centrifugation obtained in each tank cell culture autologous lymphocytes 3-5 times at a speed of 800-1200 rpm for 10 minutes for each centrifugation,

- delete after each centrifugation the supernatant and add to each a capacity of 5 ml of saline to repeat the centrifugation of the cell culture autologous lymphocytes

the fence after centrifugation of all tanks in the syringe cell mass autologous lymphocytes, adding in a syringe with typed cell mass autologous lymphocytes 10 ml of physiological solution and the introduction of the contents of the syringe intravenously to the patient in any Central or peripheral vein for 30-70 seconds

- execution of intravenous patient cell culture of autologous cells in saline solution before performing surgery, or intraoperatively, or the via 2-3 days after surgery,

- perform additional repeated intravenous patient cell culture of autologous cells in physiological solution after 7-14 days after initial injection.

Experimental studies of the proposed method of prophylaxis of purulent-inflammatory complications in trauma and orthopaedic patients using culture of autologous cells in clinical conditions showed its high efficiency. The method and its use provides a necessary and sufficient increase in the number of lymphocytes in the blood of a patient after undergoing surgery, and also provides a relatively favorable course of traumatic disease of the patient after suffering a high surgical aggression. In addition, it was ensuring the patient has the necessary increase of immunoglobulins, there is no reaction of rejection, and there have been no allergic reactions of the patient.

Implementation of the proposed method of prevention of inflammatory complications in trauma and orthopaedic patients using culture autologous lymphocytes is illustrated by the following clinical examples.

Example 1. Patient A., aged 47, arrived in 8 and orthopaedic Department FGI cyto diagnosed with the Combined tra is mA. Closed craniocerebral injury. A concussion. Closed comminuted fracture of the right femur with displacement of fragments". Analysis of the immune status showed: the total number of lymphocytes of 1.0×109/l, the absolute number of T-lymphocytes of 0.68×109/HP

The patient on day 7 after injury has performed surgery - right femur osteosynthesis plate.

Before surgical intervention performed the sampling of whole blood of the patient from his Central vein in pre-washed heparin syringe of 10 ml of Collected whole blood of the patient defended within 100 minutes. Selected lymphocytic fraction in the amount of 1.5 ml and introduced into 5 ml of culture medium RPMI-1640 and 5 ml of private patient serum. The obtained cell suspension was poured in sterile containers 2 ml and introduced into each tank 5 ml of culture medium RPMI-1640 and 0.01 ml of 10% solution of phytohemagglutinin in culture medium RPMI-1640. Containers lymphocyte culture was placed in an incubator and kept at 37°C for 72 hours. Received in each tank cell culture autologous lymphocytes were centrifuged for 5 times using a high-speed centrifuge type T-24 when the rotation speed of 800 rpm for 10 minutes each centrifugation. After each spin-on which I decanted the supernatant and in each capacity was added to 5 ml of saline to repeat the centrifugation. After centrifugation were taken from all tanks in the syringe-cell mass autologous lymphocytes, was added into the syringe with the dialed cell mass autologous lymphocytes 10 ml of physiological solution and the contents of the syringe were injected intravenously to the patient in the cubital vein of the forearm for 70 seconds, and intravenous patient cell culture of autologous cells in physiological solution was held 2 days after execution of surgical intervention. When this was implemented additional repeated intravenous patient cell culture of autologous cells in physiological solution 12 days after initial injection.

The postoperative period is smooth. Postoperative wound healing primary. Indicators of cellular immunity was growing on the next day after the introduction of the culture of autologous lymphocytes. To 3-5 days after administration of the total number of lymphocytes in the blood of the patient was 1.3×109/l, the absolute number of T lymphocytes was 0.8×109/l, the number of T-suppressors was 0,013×109/l To 10 days after administration of the total number of lymphocytes in the blood of the patient was 1.4×109/l, the absolute number of T lymphocytes was 1.0×109/l, the number of T-suppressors was 0,018×109. The definition is on the increase in the content of immunoglobulin classes G, M and A.

Sutures were removed on the 14th day. In the treatment of purulent-inflammatory complications in the patient was not observed. The fracture has healed. Allergic reactions are absent.

Example 2. Patient K., aged 65, was admitted to 8 and orthopaedic Department FGI cyto diagnosed with Right knee 3-4 degrees, severe deformity of the lower limbs, ankylosis of the right hip joint". Analysis of the immune status showed: the total number of lymphocytes of 0.95×109/l, the absolute number of T-lymphocytes of 0.64×109/HP

The patient has performed the surgery - Razmyslov osteotomy of femur and periosteal osteosynthesis of the extended lateral approach.

Before the surgery, performed the sampling of whole blood of the patient from its Central vein in pre-washed heparin syringe of 10 ml of Collected whole blood of the patient defended within 120 minutes. Selected lymphocytic fraction in the amount of 1.0 ml and introduced into 5 ml of culture medium RPMI-1640, 5 ml native serum of the patient. The obtained cell suspension was poured in sterile containers 2 ml and introduced into each tank 5 ml of culture medium RPMI-1640 and 0.01 ml of 10% solution of phytohemagglutinin in culture medium RPMI-1640. Containers lymphocyte culture were placed in thermostat and stood p and a temperature of 37°C for 72 hours. Received in each tank cell culture autologous lymphocytes were centrifuged 4 times using a high-speed centrifuge type T-24 when the rotation speed of 1200 rpm for 10 minutes each centrifugation. After each centrifugation was decanted supernatant liquid in each tank was added to 5 ml of saline to repeat the centrifugation. After centrifugation were taken from all tanks in the syringe-cell mass autologous lymphocytes, was added into the syringe with the dialed cell mass autologous lymphocytes 10 ml of physiological solution and the contents of the syringe were injected intravenously to the patient in the cubital vein of the forearm for 50 seconds, and intravenous patient cell culture of autologous cells in physiological solution was made intraoperatively.

The postoperative period is smooth. Postoperative wound healing primary. Indicators of cellular immunity was growing on the next day after the introduction of the culture of autologous lymphocytes. To 3-5 days after administration of the total number of lymphocytes in the blood of the patient was 1.25×109/l, the absolute number of T-lymphocytes was 0,75×109/l, the number of T-suppressors were 0.012×109/l To 10 days after administration of the total number of lymphocytes in the blood is aziende was 1.4×10 9/l, the absolute number of T-lymphocytes was

1,0×109/l, the number of T-suppressors was 0,017×109. Identified increased levels of immunoglobulin classes G, M, and A.

Sutures were removed on the 16th day. In the treatment of purulent-inflammatory complications, the patient was not observed. Allergic reactions are absent.

Example 3. Patient D., 69 years old, was admitted in 8 and orthopaedic Department FGI cyto diagnosed with Left knee 3-4 degrees, severe deformity of the lower limbs, ankylosis of the left hip joint". Analysis of the immune status showed: the total number of lymphocytes of 0.9×109/l, the absolute number of T-lymphocytes 0,60×109/HP

The patient has performed the surgery - Razmyslov osteotomy of femur and periosteal osteosynthesis of the extended lateral approach.

Before the surgery, performed the sampling of whole blood of the patient from its peripheral vein in a pre-rinsed with heparin syringe of 10 ml of Collected whole blood of the patient defended within 100 minutes. Selected lymphocytic fraction in the amount of 0.5 ml and introduced into 5 ml of culture medium RPMI-1640, 5 ml native serum of the patient. The obtained cell suspension was poured in sterile containers 2 ml and introduced into each tank 5 is l culture medium RPMI-1640 and 0.01 ml of 10% solution of phytohemagglutinin in culture medium RPMI-1640. Containers lymphocyte culture was placed in an incubator and kept at 37°C for 72 hours. Received in each tank cell culture autologous lymphocytes were centrifuged for 3 times using high-speed centrifuges of the type T-24 when the rotation speed of 1000 rpm for 10 minutes each centrifugation. After each centrifugation was decanted supernatant liquid in each tank was added to 5 ml of saline to repeat the centrifugation. After centrifugation were taken from all tanks in the syringe-cell mass autologous lymphocytes, was added into the syringe with the dialed cell mass autologous lymphocytes 10 ml of physiological solution and the contents of the syringe were injected intravenously to the patient in the cubital vein of the forearm for 30 seconds, and intravenous patient cell culture of autologous cells in physiological solution was carried out one day before performing surgery.

The postoperative period is smooth. Postoperative wound healing primary. Indicators of cellular immunity was growing on the next day after the introduction of the culture of autologous lymphocytes. To 3-5 days after administration of the total number of lymphocytes in the blood of the patient was 1.25×109/l, the absolute number of T-lim is acetow amounted to 0.78×10 9/l, the number of T-suppressors amounted to 0.011×109/l To 10 days after administration of the total number of lymphocytes in the blood of the patient was 1.3×109/l, the absolute number of T-lymphocytes was

0,9×109/l, the number of T-suppressors was 0,017×109. Identified increased levels of immunoglobulin classes G, M, and A.

Sutures were removed on the 14th day. In the treatment of purulent-inflammatory complications, the patient was not observed. Allergic reactions are absent.

The proposed method of prophylaxis of purulent-inflammatory complications in trauma and orthopaedic patients using culture autologous lymphocytes used in the treatment of 40 patients with different pathologies and fractures of tubular bones. Cases of purulent-inflammatory complications in patients was not observed. The fracture healed in all patients. It has been noted that the provision of strengthening the immune reactivity of patients. The proposed method does not introduce alien information, because it is based on caused by cell reactions.

1. Method of prevention of inflammatory complications in trauma and orthopaedic patients using culture autologous lymphocytes, including the blood of the patient, the cultivation of lymphocytes and intravenous their patient, distinguish the different topics the intake of whole blood of the patient is carried out before surgical intervention of any Central or peripheral vein in a pre-rinsed with heparin syringe of 10 ml and defend it within 100-140 min, select lymphocytic fraction in the amount of 0.5-1.5 ml and injected into 5 ml of culture medium RPMI-1640 and 5 ml of private patient serum is poured obtained cell suspension in sterile containers 2 ml and injected in every capacity 5 ml of culture medium RPMI-1640 and 0.01 ml of 10%aqueous solution of phytohemaglutinin in culture medium RPMI-1640, capacity, culture of lymphocytes placed in an incubator and incubated at 37°C for 72 h, and then received in each tank cell culture autologous lymphocytes centrifuged 3-5 times at a speed of 800-1200 rpm for 10 min, after each centrifugation the supernatant is poured the liquid in each tank add 5 ml of saline to repeat the centrifugation of the cell culture with autologous lymphocytes, then after centrifugation removed from all tanks in the syringe-cell mass autologous lymphocytes, in a syringe with typed cell mass autologous lymphocytes add 10 ml of saline and injected intravenous patient in any Central or perifericheskoi vein within 30-70, while intravenous patient cell mass autologous lymphocytes in the physiological solution is performed before performing surgery, or intraoperatively or within 2-3 days after surgery.

2. The method according to claim 1, characterized in that the intravenous patient cell culture of autologous cells in physiological solution additionally perform repeatedly after 7-14 days after its initial introduction.



 

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FIELD: medicine.

SUBSTANCE: sorbent for antibody removal from whole blood includes microspheres sized 0.2-1.2 mm consisting of antigen and alginic acid or calcium alginate enclosed in a semipermeable membrane made of polycationic polymer passing proteins of molecular weight 1000 kDa and less. As a polycationic polymer, the sorbent can contain either chitosan, or poly-L-lysine, or poly-L-arginine. Herewith the antigen represents a high-molecular water-soluble polymer conjugate with saccharide. Polymer represents either polyacrylamide, or polyacrylic acid, or their copolymer, or chitosan. Besides, saccharide can represent oligosaccharide of blood group A or B, or oligosaccharide component of ganglioside or sulphated glycolipid, or tumour-associated oligosaccharide. The method for making the sorbent involves injection of sodium alginate solution drops and antigen into calcium salt solution. The prepared material is kept in polycationic polymer solution to form microspheres then to be processed with complexon, e.g. ethylenediamine tetraacetate (EDTA), to remove calcium ions.

EFFECT: improved effectiveness.

6 dwg, 5 tbl, 17 ex, 10 cl

FIELD: medicine.

SUBSTANCE: therapy of the patients suffering from the hyperhomocysteinemia-accompanied diseases is ensured by a course of plasmapheresis involving removal of plasma 300-500 ml for one session. As a plasma substitute, sodium chloride 0.9% is used. The therapeutic course includes 10 sessions.

EFFECT: effective plasmapheresis therapy of diseases in such patients with considerable reduction of therapeutic complications due to substitution of great volumes of plasma removed during therapy with a solution which is not causing allergic reactions, common for the patients suffering from hyperhomocysteinemia.

3 dwg, 1 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to rheumatology, immunology and biotechnology. Substance of the invention involves procedure for reducing circulating DNA-immunocomplexes by running the blood through a prepared granulated magnet preparation with immobilised deoxyribonuclease from polyacrylamide granules characterised with magnetic properties produced by emulsion polymerisation in nitrogen gas flow.

EFFECT: advantage of the invention consists in higher sorptive performance.

3 ex, 2 tbl

FIELD: medicine; gastroenterology.

SUBSTANCE: for treatment of acute pancreatitis, arising during operative intrusions on abdominal organs, a day before operation and intraoperatively leukocytapheresis is carried out. 150 ml of leukocytal mass is taken, combined with 200 thousand units of hordox and 2.0 ml of 1% ATP solution. Mixture is incubated at temperature 24-25°C then introduced to patient by intravenous drip.

EFFECT: possibility of conservative treatment of acute pancreatitis, with no side effects due to autoleukocyte application, as well as reduction of intensive therapy terms.

1 ex

FIELD: medicine.

SUBSTANCE: invention concerns medicine, particularly hematology and obtainment of medicines out of animal and human blood. Invention claims simple method of fibrin powder obtainment, involving fibrin separation from blood clot, flushing by distilled water, drying by filter paper till complete water evaporation, and further grinding and sterilisation of target product. After drying by filter paper separated fibrin is placed for 2-4 hours to 9-10% ammonium solution in amount of 1 g of raw fibrin per 4-6 ml of solution, and then it is dried at 30-45°C till complete evaporation of water and ammonium.

EFFECT: simplified procedure, obtainment of sterile hygroscopic fibrin powder.

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