Method of determining degree of inflammatory process expression in case of knee joint osteoarthrosis

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to method of determining expression of inflammatory process in case of osteoarthrosis. Essence of method consists in the following: carried out is luminol-dependent iron-induced chemi-luminescence of model system, which has the following composition: 2.72 g of KH2PO4, 7.82 g of KCl, 1.5 g of sodium citrate C6H8O7Na3*5,5H2O per 1 liter of distilled water, pH 7.45 with 0.2 ml of 10-5 M luminol solution, after that, intensity of model system luminescence is determined in presence of synovial fluid before and after addition of synovial fluid. Degree of suppression of intensity of model system chemi-luminescence is calculated by formula. If its value is from 1.71 to 6.48%, high activity of inflammatory process is determined, if its value is from 6.49 to 21.55%, medium activity is determined, from 21.56 to 55.46 - small activity.

EFFECT: application of the method makes it possible to reduce time of determination and increases accuracy of assessment of inflammatory process degree in case of osteoarthrosis.

1 tbl, 1 dwg, 3 ex

 

The invention relates to medicine, namely to traumatology, orthopedics and rheumatology.

Osteoarthritis is the main place in the structure of the musculoskeletal system. In Russia, the average prevalence of osteoarthritis of the knee is 6.43% and reaches to 13.6% in persons older than 45 years [Ardis W. F. Prevalence of articular syndrome in Russia / W. F. Ardis, E. A. Galushko, D. I. Bazorkina, T. Y. Bolshakov // Scientific and practical rheumatology: proc. Dokl. IV Congress of rheumatology in Russia. - Kazan, 2005. - S. 146]. Diseases of the musculoskeletal system recognized socially aggravating factor, as are the second duration and the third cases of temporary disability among all classes of diseases. Osteoarthritis of the knee is the most frequent cause of chronic joint pain, and it largely determines the medical and social importance of the issue under consideration. However, early diagnosis of osteoarthritis is difficult, despite numerous studies in this field.

There is a method of determining the severity of the pathological process in osteoarthroses through the study of biochemical parameters of blood: rumorosa, antistreptolysin and C-reactive protein [Korshunov N. And., Marasai centuries, Osteoartroz // Gonarthrosis: Instrumental non-invasive methods of diagnostics is Tiki. - Yaroslavl, 2005. - S. 22-23]. However, biochemical methods usually detect pathological changes to severe stages of the disease.

It is known that in inflammatory diseases of different etiology in the blood change processes of free-radical oxidation and antioxidant activity. However, free-radical oxidation and antioxidant activity in the blood is affected by various diseases, so this indicator is not strictly specific in osteoarthritis.

The closest analogue of the invention is a method of evaluation of anti-inflammatory activity of synovial fluid in diseases of the large joints, which consists in the fact that conducting a study of chemiluminescence synovial fluid, thus determine its content of protein, then in the cuvette for chemiluminescent studies contribute intact neutrophils of healthy donors, obtained from lacokanostra, count the number of and parallel to another cell for chemiluminescent studies contribute intact neutrophils donors (NF) and synovial fluid of the patient, record the chemiluminescent response of cells of the test system, calculate the index of the chemiluminescent response (Andholdem, pulse/NF/g protein) according to the formulaAndxl= Iwith aW-IbeCwith aWNPwhere ISGthe number of pulses of the chemiluminescence response of neutrophils adding test synovial fluid, Iwithout SGthe number of pulses of the chemiluminescence response of neutrophils without adding test synovial fluid, N is the number of neutrophils donor, P - protein content (g/l) in the synovial fluid, and when the value ofholdem0.7-1.5 imp/NF/g of protein, diagnosed with moderate anti-inflammatory activity of synovial fluid, and when the value ofholdemabove 1.5 imp/NF/g protein - severe inflammatory process in the joint [patent RU 2229717, 2004]

The disadvantages of this method are the need for neutrophil donor, counting their number - indicators, which are not standardized and each time is different, as well as the determination of protein in synovial fluid of a patient, which increases the duration of the study.

The objective of the invention is to improve the efficiency of diagnosis of the severity of the inflammatory process in osteoarthritis by the study of the antioxidant activity of synovial fluid in the knee joint of the patient.

The technical result when using the invention, reducing the time and improving the accuracy of estimates of the extent of the inflammatory process in osteoarthritis of the knee.

The proposed method is as follows.

Synovial fluid taken by syringe after puncture the top of the inversion of the knee joint (recessus suprapatellaris) at the lateral edge of the base of the patella. The needle is moving perpendicular to the axis of the thigh under the tendon stretching of the quadriceps muscle to a depth of 3-4 cm and selected content. Preparing a model system of the following composition: 2,72, KH2PO4, of 7.82 g of KCL, and 1.5 g of sodium citrate C6H8O7Na3*5,5 H2O in 1 liter of distilled water. The pH of the resulting solution was adjusted to 7.45% by titration with saturated solution of KOH and add 0.2 ml of 10-5M solution lyuminola. In the cell of chemiluminometer paid 10 ml of the resulting system of standard composition, which initiate the formation of active forms of oxygen by the introduction of 1 ml of 50 mm solution of iron sulfate (FeSO4). When this develops chemiluminescence - glow accompanying the formation of free radicals. Determine its intensity for 5 min measurement. Then measure the chemiluminescence model system, which is added to 0.5 ml of the obtained patient's synovial fluid. P is the degree of inhibition of chemiluminescence model system in the presence of synovial fluid is determined by its antioxidant activity, which corresponds to the severity of the pathological process and is calculated by the formula:

(Imod-I+SYN.fluid)/Imod×100%,

where Imodthe intensity of the model system, I+SYN.fluidthe intensity of the model system after adding synovial fluid.

When the value of the degree of suppression of the intensity of chemiluminescence model system 1.71 to 6,48% define high activity of the inflammatory process, from of 6.49 to 21.55% - average activity, from 21,56 to 55,46% of small activity. The whole registration process chemiluminescence and processing results are in automatic mode, which allows to increase the accuracy and objectivity of the information received. The duration of the measurement is 5 minutes.

The invention is illustrated in the drawing, which shows the recording chemiluminescence, where 1 is a typical entry chemiluminescence model system in which adding iron salts are formed reactive oxygen species: the process is accompanied by a quick flash followed by a latent period, which goes into slow flash; 2 - after adding the synovial fluid. As the most informative measure is used as the chemiluminescence intensity during the measurement. Adding to the model system of synovial fluid inhibits chemilum is ascencio.

Under our supervision in the municipal unitary enterprise "Center of manual therapy" ,Ufa were 20 patients with osteoarthritis of the knee in age from 46 to 66 years, men and 2 women - 18. Duration of disease ranged from 1 month to 1 year. To assess the severity of the disease was determined orthopedic status that includes clinical, biochemical, instrumental, sonography, x-ray and magnetic resonance tomographic studies of the knee. The antioxidant activity of synovial fluid was determined by the degree of suppression of chemiluminescence model system, generating reactive oxygen species (table). All patients were divided into 3 groups. group 1 - patients with significant improvement after treatment, which was set low activity of the inflammatory process, group 2 - improvement with medium activity, group 3 - a slight improvement with high activity.

For the treatment of patients used non-steroidal anti-inflammatory drugs, drugs, improves microcirculation, protective agents, physiotherapy, exercise therapy, hydrokinesitherapy, massage. Intra-articular injected glucocorticosteroid drugs (betamethasone sodium phosphate, betamethasone dipropionate) intervals through the day. The total number of injections ranged from 2 to 3 treatments. After copyrow is of synovitis weekly was introduced inside the joint "Hyalgan Fidia" from 1 to 3 injections per course of treatment.

All patients (n=20) in the process of treatment, the degree of suppression of chemiluminescence model system adding synovial fluid is increased, which indicated the increase of its antioxidant activity.

Clinical example 1

The patient 3. 1947 R., appealed to the municipal unitary enterprise "WTC" with complaints of pain in the left knee joint, limitation of movements.

Status localis: During the inspection region of the left knee revealed swelling, palpation in the projection of the joint space - pain. Motion limited. Bend the left knee joint 180-75 degrees, there limp on the left leg. While the survey found: complete blood count (ESR 48 mm/hour), ultrasound (in the cavities of the left knee moderate amount of inhomogeneous fluids, predominantly in the upper inversion and lateral inversions, articular gap narrowed) and x-ray of the left knee (signs of deforming arthrosis of the I century).

Diagnosis: Osteoarthritis of the left knee of the I century Synovitis. A survey of the proposed method. Imod- 42,27, I+SYN.- 20,22 degree of suppression of the intensity of chemiluminescence model system was 52,16, which indicates a low activity of the inflammatory process.

After out-patient treatment no complaints, WM is avnoj syndrome was arrested.

Status localis: When viewed from the left knee joint swelling was not detected, palpation in the projection of the joint space painless. Bend the left knee joint 180-125 degrees. Walking time restored.

The study of the antioxidant activity of synovial fluid in the knee joints showed an increase. Significant improvement of the condition, the severity of the inflammatory process is minimal.

Clinical example 2

Patient S., 1966 R., appealed to the municipal unitary enterprise "WTC" with complaints of pain in the knee joints, limitation of movement and discomfort.

Status localis: Upon examination of the knee joints revealed swelling, contours smoothed palpation in the projection of the joint space soreness. Motion limited. Bend the left knee joint 180-75 degrees. The bend in the right knee joint 180-80 degrees, there limp on the right leg.

While the survey found: complete blood count (erythrocyte sedimentation rate of 19 mm/hour), radiography of the knee joints (signs of deforming arthrosis II century).

Diagnosis: Bilateral gonarthrosis II senior Synovitis.

A survey of the proposed method. Imod- 14,64, I+SYN.fluid- 12,30 degree of suppression of the intensity of chemiluminescence model system was 15,98, which indicates the average activity populationincrease.

After out-patient treatment no complaints, articular syndrome is under control.

Status localis: When viewed in the area of knee joints swelling is not revealed, palpation in the projection of the joint space painless. Flex at the knee joints 180-120 degrees. Walking time restored. During the treatment the antioxidant activity of synovial fluid increased, the intensity of the inflammatory process is decreased, the marked improvement of the patient.

Clinical example 3

Patient O., 1951 R., appealed to the municipal unitary enterprise "WTC" with complaints of pain in the right knee joint, limitation of movements.

Status localis: When inspecting the area of the right knee revealed swelling, palpation in the projection of the joint space and the displacement of the patella, pain. Motion limited. The bend in the right knee joint 180-80 degrees, there limp on the right leg.

While the survey found: complete blood count (erythrocyte sedimentation rate of 17 mm/h), ultrasound examination of the right knee joint (determined thinning of the cartilage surface, the phenomenon of synovitis, degenerative-dystrophic changes in the anterior horn of the medial meniscus. Echo signs of gonarthrosis Art. II, synovitis).

Diagnosis: Octoarts right knee II senior Synovitis.

A survey of the proposed method. Imod- 7,84, I+SYN.- 26,69 degree of suppression of the intensity of chemiluminescence model system was 4,13, which indicates a high activity of the inflammatory process.

After out-patient treatment no complaints, pain syndrome was arrested.

Status localis: When viewed in the right knee joint swelling was not detected, palpation in the projection of the joint space painless. The bend in the right knee joint 180-115 degrees.

The patient's condition has improved slightly. The antioxidant activity of synovial fluid has not changed.

The method of determining the severity of the inflammatory process in the osteartrozis, including chemiluminescent study using synovial fluid, characterized in that conduct Lomonosovsky gelatination the chemiluminescence model system, which has the following composition: 2,72 g KH2PO4, of 7.82 g of KCL, and 1.5 g of sodium citrate C6H8O7Na3*5,5 H2O in 1 liter distilled water, pH was 7.45 with 0.2 ml of 10-5M solution lyuminola, then in the presence of synovial fluid determine the intensity of the glow model system before and after adding the synovial fluid, calculate the degree of suppression of the intensity of chemiluminescence model system by the formula:
(Imod-I +SYN.fluid)/Imod×100%
where
Imodthe intensity of the model system,
I+SYN.fluidthe intensity of the model system after adding synovial fluid,
and when it is 1.71 to 6,48% define high activity of the inflammatory process, from of 6.49 to 21.55% - average activity, from 21,56 to 55,46% of small activity.



 

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2 tbl, 3 ex

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2 ex

FIELD: medicine, psychiatry.

SUBSTANCE: one should isolate DNA out of lymphocytes of peripheral venous blood, then due to the method of polymerase chain reaction of DNA synthesis one should amplify the fragments of hSERT locus of serotonin carrier gene and at detecting genotype 12/10 one should predict the risk for the development of hallucino-delirious forms of psychoses of cerebro-atherosclerotic genesis.

EFFECT: more objective prediction of disease development.

3 ex

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