Erythrocyte diagnosticum and a method for the diagnosis of chronic intoxication by inorganic vanadium compounds

 

(57) Abstract:

Erythrocyte diagnosticum and a method for the diagnosis of chronic occupational poisoning with inorganic compounds of vanadium relates to the field of medicine, namely to the laboratory diagnosis of occupational diseases. The essence of the claimed group of inventions is that the connection of the leukocytes of a patient with an equal volume of a 0.5% suspension patriciavonne sheep erythrocytes loaded with inorganic compounds of vanadium, incubated for 10-15 minutes at a temperature of up 37.0-37.5 C, centrifuged, do a PAP smear, fix it, dried, stained by Romanovsky-Institute. Looking at under the microscope 100 lymphocytes, noting among these cells, committed by three or more sheep erythrocytes (ROCK). When the number of ROCK-specific inorganic compounds of vanadium compared to the norm, diagnose the presence of inorganic compounds of vanadium in the body and diagnose: professional intoxication higher aliphatic fatty amines series. The technical result is to increase the objectivity and accuracy of the diagnosis of occupational intoxication by inorganic compounds, of vanadium. 2 C. p. F.-ly, x disease, and can be used for diagnosis of chronic occupational poisoning with inorganic compounds of vanadium.

The purpose of the invention is to increase the objectivity and accuracy of diagnosis of occupational diseases caused by inorganic compounds, of vanadium.

The prototype of the present invention is a Method for the diagnosis of intoxication higher aliphatic fatty amines number” [T. A. Yushkova in. A. Yushkov, N. With.Sedinina //AU No. 1783438, BI No. 47], which are as follows: connect 0.1 ml leukocytes of a patient with an equal volume of a 0.5% suspension patriciavonne sheep erythrocytes loaded WEAR. The mixture was incubated for 10-15 minutes at a temperature of up 37.0-37.5 C, centrifuged, do a PAP smear, fix it, dried, stained by Romanovsky-Institute. Looking at under the microscope 100 lymphocytes, noting among these cells, committed by three or more sheep erythrocytes (ROCK). When the number of ROCK-specific WEAR, compared with the norm (3,40,5%) diagnose the presence WEAR in the body and diagnose: professional intoxication higher aliphatic fatty amines number.

The proposed invention differs from the prototype in that the establishment is).

Original erythrocyte diagnosticum specific inorganic compounds of vanadium, prepared as follows.

For sensitization of erythrocytes prepare a solution with a concentration of Vanadate sodium 0.1 mg/ml in buffered physiological solution of pH 6.4. 1 ml of 5% suspension patriciavonne erythrocytes centrifuged, the supernatant removed, and the precipitate erythrocytes poured 1 ml of a solution of sodium Vanadate. The mixture was thoroughly shaken and incubated at room temperature for 25-30 minutes, after which the red blood cells washed twice by centrifugation at 1500 rpm for 10 min isotonic sodium chloride solution and they lead up to a 0.5% suspension.

In the intact laboratory animals (rats) the number of ROCK, the specific vanadium, is 5.3±0,8% (table.1), and 30 minutes after intraperitoneal administration of Vanadate sodium at a dose of 1 MPC -38,21,1% (p1<0,001) or vanadium pentoxide at a dose of 1 MPC -34,0±2.3% (p1<0,001; p2>0,05).

The specificity of diagnosticum proved by treatment with excess sodium Vanadate or WEAR lymphocytes obtained from rats exposed to the sodium Vanadate. If in the first case the level of the ROCK was significantly decreased and was equal to what viewlanguage effect of sodium Vanadate.

The essence of the invention is the detection of inorganic compounds of vanadium based on the ability of lymphocytes of the body to sencibilisiruet to the substance falls into the body in any way that is accompanied by activation in the number of receptors for these compounds vanadium.

The essence of the method consists in determining the number of lymphocytes that can interact with sheep erythrocytes sensitized inorganic vanadium.

The method is as follows. Take 0.1-0.2 ml of patient's blood in 0.1-0.2 ml of a 5% solution of sodium citrate to prevent clotting. After thorough shaking the tubes in it add to lizirovania erythrocytes 1 ml of distilled water for 30 sec, and then poured 1 ml of 1.8% sodium chloride solution to restore pH 7.0. The tubes centrifuged for 5-10 min at 1500 rpm, after which the supernatant liquid is drained and the sediment leukocytes resuspended in 0.1-0.2 ml isotonic sodium chloride or medium 199. To the resulting suspension add 0.1-0.2 ml of 0.5% suspension patriciavonne sheep erythrocytes loaded with inorganic compounds of vanadium, the mixture is incubated for 10-15 min at t 37,0-37.5 C, then centrtekstil serum (bovine, horse, human). From the obtained cell suspension do a PAP smear, fixed in methyl alcohol or ethyl alcohol, dried, stained by Romanovsky-Institute. Under the microscope consider 100 lymphocytes, noting among them are the number (%) of cells, committed by three or more erythrocytes (ROCK).

Processing of sheep erythrocytes in glutaric the aldehyde is produced as follows: combine 1 ml of 5% suspension thoroughly washed native sheep red blood cells with 0.25% solution of glutaric aldehyde in the ratio of 1:1, the mixture is maintained at 37,0 C for 15 min, after which the red blood cells washed twice from aldehyde buffered isotonic sodium chloride pH 7.2 and brought to the original volume of the same solution (5% suspension).

Control the “loading” of erythrocytes vanadium was determined by atomic emission method additives on diffraction spectrograph DFS-452 with a universal electronic generator UGE-4 (PL.2).

When using atomic emissionnogo discovery of vanadium found that vanadium detected on erythrocytes loaded with sodium Vanadate, but not on intact (control) erythrocytes (see tab.3).

Examples of practical use of the method.

Example 1. B-e a-VA, 32, apparatchick shop ferrovanadium. Work experience of 10 years.

Complaints of headaches, dizziness, epigastric pain, flatulence, menstrual disorders.

U b-s was taken in a test tube blood 0,1 ml 0,1 ml of 5% sodium citrate, thoroughly mixed for lysis of erythrocytes for 30 seconds, was added 1 ml of distilled water, and then to restore pH 7.0 was added 1 ml of 1.8 % isotonic sodium chloride. The tube was centrifuged for 5-10 min at 1500 rpm, after which adosados decanted, the precipitate resuspendable in 0.1 ml of isotonic sodium chloride to the suspension was added 0.1 ml of a 0.5% suspension of erythrocytes loaded with sodium Vanadate. The mixture was incubated for 10-15 min at t=37-37,5 C, and then centrifuged for 5-10 min at 1500 rpm the Supernatant was aspirated with a pipette, and the residue resuspendable in the drop intact horse serum. On a clean glass slide suffered a drop of the suspension of the tubes, made a smear, dried it, fixed in ethanol, stained by Romanovsky-Institute. Smear has microscopically under immersion (90), counted 100 cells, noting the account in a b-s A-howling found 26% of the ROCK, the specific sodium Vanadate, whereas normal their level equal 5,90,6%.

Diagnosis: Chronic occupational poisoning with inorganic compounds of vanadium; the imbalance of the hypothalamic-pituitary-adrenal system and, as a consequence, ovarian dysfunction, impaired estrogen-androgen balance, premenstrual syndrome, chronic gastroenterocolitis, pyelonephritis.

Example 2. B-Nai E-WA, 48, apparatchick shop ferrovanadium. Work experience 26 years.

Complaints of weakness, fatigue, headaches, dizziness, shortness of breath, pain in the right hypochondrium and epigastrium, disruption of the menstrual cycle.

U b-s was taken in a test tube blood 0,1 ml 0,1 ml of 5% sodium citrate, thoroughly mixed for lysis of erythrocytes for 30 seconds, was added 1 ml of distilled water, and then to restore pH 7.0 was added 1 ml of 1.8 % isotonic sodium chloride. The tube was centrifuged for 5-10 minutes at 1500 rpm, after which adosados decanted, the precipitate resuspendable in 0.1 ml of isotonic sodium chloride to the suspension was added 0.1 ml of a 0.5% suspension of erythrocytes loaded with sodium Vanadate. The mixture was incubated for 10-15 min at t=37-37,5 C, and then centrifuged for 5-10 min th serum. On a clean glass slide suffered a drop of the suspension of the tubes, made a smear, dried it, fixed in ethanol, stained by Romanovsky-Institute. Smear has microscopically under immersion (90), counted 100 lymphocytes, noting among them are the number (%) of outlets (ROCK), i.e., cells, committed on its surface three or more of the erythrocyte. When counting in a b-s E-howl found 39% of the ROCK, the specific sodium Vanadate, whereas normal their level equal 5,90,6%.

Diagnosis: Chronic occupational poisoning with inorganic compounds of vanadium; chronic gastroenterocolitis, oppression of microsomal enzymes, fermentopathy, suppression of the hypothalamic-pituitary-adrenal system and, as a consequence, ovarian dysfunction, impaired estrogen-androgen balance, the debut of the climacteric syndrome; chronic pyelonephritis.

Example 3. B-Neu St-n, 47 years old, apparatchik shop ferrovanadium. Experience 25 years.

Complaints: weakness, fatigue, headaches, dizziness, shortness of breath, mild irritability, excessive sweating, low-grade fever, pain in the epigastrium and right hypochondrium and lower potency.

U b-s took in tube shelter is authorized water and then to restore pH 7.0 was added 1 ml of 1.8 % isotonic sodium chloride. The tube was centrifuged for 5-10 min at 1500 rpm, after which adosados decanted, the precipitate resuspendable in 0.1 ml of isotonic sodium chloride to the suspension was added 0.1 ml of a 0.5% suspension of erythrocytes loaded with sodium Vanadate. The mixture was incubated for 10-15 min at t=37-37,5 C, and then centrifuged for 5-10 min at 1500 rpm the Supernatant was aspirated with a pipette, and the residue resuspendable in the drop intact horse serum. On a clean glass slide suffered a drop of the suspension of the tubes, made a smear, dried it, fixed in ethanol, stained by Romanovsky-Institute. Smear has microscopically under immersion (90), counted 100 lymphocytes, noting among them are the number (%) of outlets (ROCK), i.e., cells, committed on its surface three or more of the erythrocyte. When counting in a b-main St-found on 43% of the ROCK, the specific sodium Vanadate, whereas normal their level equal 5,90,6%.

Diagnosis: Chronic occupational poisoning with inorganic compounds of vanadium; the imbalance of the hypothalamic-pituitary-adrenal system, chronic prostatitis, violation of diuresis, chronic peel stickum, specific inorganic compounds of vanadium, namely, that for sensitization of erythrocytes prepare a solution with a concentration of Vanadate sodium 0.1 mg/ml in buffered saline solution pH 6.4, which is combined with 1 ml of 5% suspension patriciavonne erythrocytes, the mixture was thoroughly shaken and incubated at room temperature for 25-30 minutes, after which the red blood cells washed twice by centrifugation at 1500 rpm for 10 min isotonic sodium chloride solution and they lead up to a 0.5% suspension.

2. The way to diagnose the erythrocyte diagnosticum obtained under item 1, intoxication vanadium by examination of the patient, characterized in that the blood leukocytes of the patient add an equal volume of a 0.5% suspension patriciavonne sheep erythrocytes loaded with sodium Vanadate, the mixture is incubated at 37-37,5 C for 10-15 min, centrifuged at 1500 rpm for 5-10 min, the supernatant removed with a pipette, sediment resuspended in the drop of any intact serum: (bovine, horse, human), do a PAP smear, fixed in methyl alcohol or ethyl alcohol, dried, stained by Romanovsky-Institute, count in a number of rosette lymphocytes and increased the organic compounds of vanadium.

 

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