RU2545805C2 - Method of early diagnostics of fat embolism syndrome in case of fractures of long tubular bones and pelvis bones - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to traumatology and orthopaedics; can be used for the early diagnostics of fat embolism syndrome in case of fractures of long tubular bones and pelvis bones and control of performed treatment efficiency. The method of fat embolism syndrome diagnostics consists in the analysis of serum of venous blood, sampled from a patient on the first day of the post-traumatic period, and determination of the S100B protein concentration in it by enzyme immunoassay. If the protein S100B norm, constituting 125 ng/l, is exceeded, the fat embolism syndrome is diagnosed.

EFFECT: method makes it possible to increase the accuracy of early FES without the necessity to perform spinal puncture or femoral artery puncture, which can result in serious complications.

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Description

The invention relates to medicine, namely to traumatology and orthopedics, and for the early diagnosis of fat embolism syndrome (SJE) for fractures of long tubular bones and pelvic bones and monitoring the effectiveness of the treatment.

According to statistics, more than 5 million people die from injuries in the world every year, in Russia - more than 300 thousand. From severe complications of a traumatic disease, 15-20% of all victims with severe combined trauma die. One of these complications is SJE.

SZHE can be defined as a clinical condition characterized by impaired lung and central nervous system functions due to obstruction of microvessels by large globules of fat, occurring mainly after severe injuries with fractures of long tubular or pelvic bones (Chechetkin A.V., Tsybulyak G.N. Infusion Transfusion Therapy for Fat Embolism Syndrome Transfusiology, 2003 (2): p. 42-51).

There is a method for diagnosing the cerebral form of traumatic embolism, when a spinal cerebrospinal fluid is taken from a patient, fat globules stained with Sudan IV solution are measured in the first 10-15 minutes under a microscope and, in the presence of round drops of red or bright pink color, the cerebral form of traumatic emboli is diagnosed ( RF patent No. 2176798, IPC G01N 33/92, 2000).

The disadvantage of this method is the need for spinal puncture, which can be complicated by neurological complications (neuropathy, spinal cord injury). In addition, changes are already detected with a developed clinical picture and, as a rule, only confirm the clinical diagnosis.

Certain diagnostic value of magnetic resonance imaging of the brain is also known, which can identify areas of high intensity T2 weighted images (Takahashi, Μ. Et al. Magnetic resonance imaging findings in cerebral fat embolism: correlation with clinical manifestations. J. Trauma, 1999. 46 (2): p. 324-327). MPT is useful for patients with neurological symptoms of fat embolism.

The closest in technical essence is a method for the diagnosis of traumatic fat embolism, which is carried out by determining the patient's blood serum under the microscope of stained fat globules. In this case, blood for analysis is taken from the femoral artery and central vein, blood serum is applied directly to a glass slide, a Sudan solution is added, and after 1 min microscope (RF patent No. 2195659, IPC G01N 33/483, 2000).

The disadvantage of this method is the need for puncture of the femoral artery and central vein, in which there is a high risk of infection, thrombosis, hematomas and the occurrence of false aneurysms. Using this method, fat globulinemia is diagnosed, which is detected in 95% of patients with fractures, but cannot diagnose SJE.

The objective of the present invention is to remedy these disadvantages, namely improving the accuracy of early diagnosis of SLE without the need for spinal puncture or puncture of the femoral artery, which can lead to serious complications, and the fact that the method does not require expensive equipment, such as a magnetic resonance imager.

This problem is solved by the fact that in the method for early diagnosis of fat embolism syndrome in fractures of long tubular bones and pelvic bones, including preparation of blood serum, blood is taken from the cubital vein on the first day of the post-traumatic period and its subsequent centrifugation, the protein concentration S100B is determined in the obtained serum enzyme-linked immunosorbent assay non-competitive analysis and with an increase in the norm of the S100B protein of 125 ng / l, the presence of fat embolism syndrome is diagnosed.

It is known that the neuroglial protein S100B, a calcium-binding protein, is produced and secreted mainly by glial and Schwann cells of the central nervous system.

When implementing this method, enzyme-linked immunosorbent assay non-competitive analysis was based on the use of two types of murine monoclonal antibodies that specifically recognize two different epitopes of the S100B molecule - S100A1B and S100BB.

In the analysis of the S100B protein, good results were achieved, for example, using the CanAg S100 диагност diagnostic kit manufactured in Sweden. Standard solutions, a solution of conjugate labeled with horseradish peroxidase, wash buffer are prepared according to the instructions for the set. Patient sera standards and samples are incubated with biotinylated monoclonal antibodies to S100B (MAb) S23 in streptavidin-coated microplate cells. During the incubation process, S100B, present in samples and standards, is adsorbed in streptavidin-coated wells due to specific binding to biotinylated antibodies. The wells are then washed and incubated with monoclonal antibodies to S100B (MAb) S53 labeled with horseradish peroxidase (HRP). After washing, a buffered substrate / chromogen solution (hydrogen peroxide and 3,3 ′, 5,5′-tetramethylbenzidine) is added to each well, as a result of which an enzymatic reaction occurs. During the reaction, a blue color appears in the presence of antigen. The color intensity is proportional to the amount of S100B present in the sample. The color intensity is measured on a microplate reader with a filter of 620 nm (at 405 nm after adding a stop solution). A plot of the optical density versus protein concentration is plotted for each analysis and the concentration of S100B in the patient’s samples is calculated from it. Express the level of protein S100B in the serum of the patient's venous blood in ng / L.

The upper limit of normal values was proposed to consider the concentration of S100B protein at 125 ng / L. The normal value of S100B protein in serum of venous blood was established based on an analysis of the data described in the sources of scientific and technical information (Muller, K., et al. S100B serum level predicts computed tomography findings after minor head injury. J. Trauma, 2007. 62 (6): p. 1452-1456; Sorokina, E.G. et al. Protein S100B and its autoantibodies in the diagnosis of brain injuries in traumatic brain injuries in children. Journal of Neurology and Psychiatry, 2010 (8): p. 30 -35) and specified in the standards by the developer, as well as on the basis of our own studies obtained during the examination of donors.

Own research data are presented in table 1.

Table 1 Concentrations of S100B Protein in Serum of Venous Blood of Patients in the Early Post-Traumatic Period Post-traumatic period The concentration of protein S100B, ng / l groups the control I (fracture) II (SZHE) 1 day 66.33 ± 11.19 79.72 ± 7.54 229.77 ± 51.43 * 2 days 40.50 ± 10.10 60.72 ± 11.90 113.90 ± 24.99 3 days 52.50 ± 2.80 63.60 ± 11.02 230.13 ± 108.87 Note: Group I (fracture): 5 patients with polytrauma without established fat embolism syndrome. Group II (SJE): 3 patients with established fat embolism syndrome (according to Gurd) [21]. Control Group: 3 healthy volunteers. Groups were comparable in gender and age, but differed in severity * - the difference is significant, p <0.05 (Kruskal-Wallis test)

Examples of specific performance.

Example 1. Patient S., 74 years old, was hospitalized after a road injury with a diagnosis of polytrauma. Open displaced comminuted fracture of both bones cf / 3 of the right tibia. Closed biased fracture of the middle third of the right femur. Closed biased fracture of the middle third of the left femur. Abrasions of the right lower leg. Traumatic shock II Art. According to emergency indications performed:

1. Primary surgical treatment of a wound. 2. Osteosynthesis of the right femur with the Ilizarov rod apparatus. 3. Osteosynthesis of the left femur with the Ilizarov rod apparatus. 4. Osteosynthesis of bones of the right lower leg with the Ilizarov spoke device. On the second day, the patient developed encephalopathy, respiratory failure began to increase. Traumatic fat embolism is suspected. The patient underwent a blood test for fat globules. The III degree of fat globulinemia is defined. The level of S100B protein in the patient in the blood was determined in the first day of the post-traumatic period. The result is 332.4 ng / L.

This symptomatology allowed the patient to be diagnosed with a cerebral form of fat embolism.

The patient was treated in the intensive care unit. Artificial ventilation of the lungs, infusion, transfusion therapy, antibiotics, and blood rheology improving agents were performed. Subsequently, the patient was discharged from the hospital in satisfactory condition.

Example 2. Patient S., 50 years old, was hospitalized after a road injury with a diagnosis of multiple injury. Closed fracture of the right pubic and sciatic bones with a slight displacement. Open comminuted fracture of bones of the right tibia. An open comminuted fracture of the right humerus with displacement. Closed fracture of the acromial end of the right clavicle with displacement. Dumb abdominal injury. Dull chest injury. Closed fracture 3, 4, 7, 8 ribs on the right, 9 ribs on the left. Hemopneumothorax on the right. Torn wounds of the nose, cf / 3 of the right lower leg, right hand, right shoulder joint. Traumatic shock II Art. The patient was hospitalized in ICU. The next day the patient developed severe encephalopathy, respiratory failure. The patient underwent a blood test for fat globules. The IV degree of fat globulinemia is determined. In a patient, the S100B protein was determined in the blood on the first day of the post-traumatic period. The result is 184.4 ng / L.

Diagnosed with Fat Embolism Syndrome. Conducted antishock therapy, infusion, multiple blood transfusions, analgesics, antibiotics. In order to stabilize the condition, the following was operated: an extrafocal PSC according to Ilizarov of the right lower leg with a 4-section apparatus. After preoperative antibacterial prophylaxis. After stabilization, the patient was transferred to the trauma unit. The wounds healed by primary intention. The seams are removed. Discharged in satisfactory condition.

The proposed method for the diagnosis of fat embolism syndrome does not cause complications, is a reliable, relatively inexpensive and simple method based on modern achievements in the field of metabolomics. It allows the first day to identify SZHE with skeletal injury and to monitor the effectiveness of its treatment.

Information sources

1. About some features of injuries in the Russian Federation / S.A. Leonov, E.V. Ogryzko, N.M. Zaichenko. - FGU TSNIIOIZ Roszdrav, Moscow. http://vestnik.mednet.ru/content/view/143/30/lang.ru/.-

2. Abbott, M.G., Fat embolism syndrome: An in-depth review. Asian Journal of Critical Care, 2005 (1): p. 19-24.

3. Chechetkin, A.B., Tsybulyak G.N. Infusion-transfusion therapy for fat embolism syndrome. Transfusiology, 2003 (2): p. 42-51.

4. RU 2176798 C2, publ. 12/10/2001.

5. Takahashi, M. et al. Magnetic resonance imaging findings in cerebral fat embolism: correlation with clinical manifestations. J Trauma, 1999.46 (2): p. 324-7.

6. RU 2195659 C2, publ. 12/27/2002.

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9. Donato, R. S100: a multigenic family of calcium-modulated proteins of the EF-hand type with intracellular and extracellular functional roles. Int J Biochem Cell Biol, 2001.33 (7): p. 637-68.

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12. Ingebrigtsen, T. et al. Traumatic brain damage in minor head injury: relation of serum S-100 protein measurements to magnetic resonance imaging and neurobehavioral outcome. Neurosurgery, 1999.45 (3): p. 468-75; discussion 475-6.

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Claims (1)

A method for early diagnosis of fat embolism syndrome in fractures of long tubular and pelvic bones, including the preparation of blood serum, characterized in that on the first day of the post-traumatic period, blood is taken from the cubital vein and its subsequent centrifugation, the concentration of protein S100B is determined by enzyme-linked immunosorbent assay and with an increase in the norm of S100B protein of 125 ng / l, the presence of fat embolism syndrome is diagnosed.