RU2178563C1 - Method for predicting premorbid state under exposure to chemical pollutants in production environment - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: method involves taking venous blood and urine samples in the amount of 10 ml in the empty stomach in the morning. 0.5 ml of taken plasma (serum) is diluted with 18.5 ml of phosphate buffer solution having pH of 7.45. Chemiluminescence is induced by adding 1 ml of 50 mMole FeSO₄•7H₂O solution and measurement are taken during 10 min. Spontaneous luminescence and chemiluminescence optic sum are calculated from the moment of rash flash end to slow flash beginning. Spontaneous blood luminescence being equal to or less than 2,0•10⁵ quanta/min and greater than 7,0•10⁵ quanta/min, luminescence optic sum being equal to or less than 10,0•10⁵ quanta/min and equal to or greater than 40,0•10⁵ quanta/min and in urine spontaneous luminescence being equal to 6,0•10⁵ quanta/min or less and 12,0•10⁵ quanta/min and greater, luminescence optic sum being equal to or less than 16,0•10⁵ and equal to and greater than 70,0•10⁵ quanta/min, protective adaptation mechanism disorders are considered to take place and premorbid state development is predicted to occur. EFFECT: high accuracy and reliability of the method; simplified and accelerated examination procedure. 10 dwg, 2 tbl

Description

 The invention relates to medicine, in particular to medical ecology, hygiene, toxicology, occupational pathology, can be used to predict premorbid state, namely for early detection of violations of adaptive-adaptive reactions of the body, diagnosis of prenosological conditions, prediction of pathological changes when exposed to chemical factors production environment.

 The literature describes methods for differential diagnosis of the state of a whole organism under the influence of adverse factors that allow determining the status of the organism as a norm, a borderline state between norm and pathology, pathology (Mamedov T.G., 1982; Mukhambetova L. X., 1997; Piruzyan L. . A. et al., 2000; Serdyukovskaya G.N., Churyanova N.I. 1995; Sidorenko G.N., Merkuryeva R.V. 1991).

The prototype of the invention is a method of non-invasive biochemical diagnosis of the state of public health (Mukhambetova L. X., 1997), which proposed a methodological scheme for biochemical analysis of available human biomaterial, including the determination of various biomarkers (enzymes, biosubstrates, metabolites of endogenous and exogenous origin, xenobiotics). "Non-invasive biochemical diagnosis of the state of health of the population of Moscow." In the book. : Hygienic assessment of the environment and public health in Moscow. - M., 1997 .-- S. 41-44. The proposed method allows to assess the state of detoxification processes and the removal of toxic products from the body, the barrier functions of biomembranes, the activity of the main vital processes (energy supply, catabolism, etc.). The following urine biomarkers are determined:
- enzymes (lactate dehydrogenase, glutathione transferase, acetyl esterase, β-glucuronidase, N-acetyl β-D-glucosaminidase, β-galactosidase, phenolsulfotransferase, etc.);
- trace elements (calcium, sodium, potassium, etc.);
- creatinine, uric acid;
- coproporphyrins (δ-amino-levulinic acid, etc.);
- metabolites (endogenous and exogenous);
- protein according to Lowry; -
- the state of cytochrome P ₄₅₀ -dependent monooxygenase system;
- the content of lipid peroxidation products (diene conjugates, malondialdehyde, Schiff base)
The use of non-invasive biochemical diagnostics reveals among the examined individuals a group of practically healthy people who did not have significant biochemical deviations at the time of the examination, a group with unstable deviations of the studied parameters, and a group with stable and significantly pronounced changes in several (at least 2) indicators, which makes it possible to distinguish it as a risk group for the development of pathologies of a chemical nature. However, this method is very time-consuming, requires a large number of hard-to-reach, commercially expensive reagents, consists of many sequential steps (cooling, incubation, temperature control, centrifugation, etc.), is time-consuming, which severely limits its use in mass clinical examinations and preventive examinations .

 The described approach indicates a violation of the metabolic processes of the functioning of the kidneys.

 The technical result is to increase the accuracy and reliability of the method, simplifying the study, as well as reducing time.

 In FIG. 1 and 2 shows a record of chemiluminescence (CL) of urine and blood according to example 1.

 In FIG. 3 and 4 - record CL urine and blood according to example 2.

 In FIG. 5 and 6 - recording CL urine and blood according to example 3.

 In FIG. 7 and 8 - recording CL urine and blood according to example 4.

 In FIG. 9 and 10 - record CL urine and blood according to example 5.

The proposed method is as follows: blood from a vein in an amount of 10 ml is taken in the morning on an empty stomach. Serum is obtained after removal of the formed clot. Plasma is prepared using sodium citrate (5 mg / ml) as an anticoagulant. The shaped elements are precipitated by centrifugation. The resulting plasma and blood serum is stored at a temperature of +4 ^o C for no more than 8 hours. 0.5 ml of plasma (serum) is diluted in 18.5 ml of saline solution of the following content: 20 mm KH ₂ PO ₄ , 105 mm KCl, pH 7.45. Chemiluminescence is initiated by the addition of 1 ml of FeSO ₄ • 7H ₂ O, the optimal concentration of which is 2.5 mM in the incubation medium.

Urine for the study of chemiluminescence is taken in the morning, its pH is measured and, if necessary, adjusted to 5.0-6.0 units. The sample volume is 10 ml. Chemiluminescence is initiated by the addition of an iron sulfate salt (1 ml of FeSO ₄ • 7H ₂ O — 50 mM) and measured over 10 minutes.

 The intensity of CL is recorded on the device HLM-003, included in the register of equipment manufactured in Russia.

The sequence of operations when measuring CL was as follows. In the thermostatic cuvette of the device equipped with a stirrer, the test samples are placed. After mixing and equalizing the temperature of the contents of the cuvette chamber, the dark current is measured for one minute, then the shutter of the device is opened and the intensity of the spontaneous luminescence induced by the addition of Fe ^{+ 2} salts is recorded. At the moment the luminescence reaches the stationary level, when the accumulation of peroxide products ceases and the main part of Fe ^{+ 2} ions is already used up, the experiment is completed and the shutter of the device is closed. A check of the stability of the installation is carried out before each measurement by radiation of the secondary standard SFKhM-1 (GOST-9411-81). The luminous intensity of the control sample is 5.1 • 10 ⁵ quanta per second. For convenience, this value is taken as one relative unit (rel. Units). The value of the spontaneous luminescence, the intensity of the fast flash at the moment the initiator is added, the duration of the latent period, the steepness of the rise of the slow flash, and the CL light sum for 5 minutes of recording are determined (for blood and urine).

 - The spontaneous glow determines the speed of the SRO without external intervention.

 - A quick flash occurs when an oxidation initiator is added. The flash amplitude is directly proportional to the content of peroxide products.

 - The induction period or latent period characterizes antioxidant properties.

 - The steepness of the growth of a slow flash is determined by the rate of initiation of SRO.

 - The maximum amplitude of the slow flash and the light sum of the glow characterize the ability of the biological material to undergo oxidation.

For reliable prediction, a single study of blood and urine is sufficient. With spontaneous luminescence (Cn) of blood 2.0 • 10 ⁵ quanta / min and below and 7.0 • 10 ⁵ quanta / min and above, the luminosity sum (S) - 10.0 • 10 ⁵ quanta / min and below and 40 , 0 • 10 ⁵ quanta / min and above, and in urine - Cn - 6.0 • 10 ⁵ quanta / min and below and 12.0 • 10 ⁵ quanta / min and above, S - 16.0 • 10 ⁵ quanta / min and lower and 70.0 • 10 ⁵ quanta / min and higher predict the development of premorbid state. Other parameters of chemiluminescence - the amplitude of the fast flash (A) and the maximum amplitude of the slow flash (Max) do not provide significant additional information on the change in the light sum of the glow (Tables 1 and 2).

 A clinical examination of 860 workers in petrochemical industries was conducted (OAO Ufaneftekhim, OAO Ufakhimprom, OAO Salavatnefteorgsintez, ZAO Sterlitamaksky Petrochemical Plant). During the year, 486 of the examined individuals underwent a medical and health-improving course at the sanatorium on-the-job on-site basis (OAO Ufaneftekhim, OAO Ufakhimprom, OAO Salavatnefteorgsintez, ZAO Sterlitamaksky Petrochemical Plant).

 Upon admission, during treatment and before discharge, the chemiluminescence of blood and urine was examined. In 22.3% of people sent to the sanatorium, the CL and blood urine intensity at the time of admission was higher than normal, in 15.2% it was lower than the control values, i.e. this was more than 1/3 of the examined.

 Example 1

 Sitdikov R. M., 42 years old, works as an operator in OAO Ufaneftekhim.

Complete blood count: hemoglobin 135 g / l, red blood cells 4.5 • 10 ¹² / l, color indicator (CP) 0.9, white blood cells 8.2 • 10 ⁹ / l, erythrocyte sedimentation rate (ESR) 16 mm / hour, platelets 170 • 10 ⁹ / l, stab-type 0.3 • 10 ⁹ / l, segmented 4.5 • 10 ⁹ / l, eosinophils 0.02 • 10 ⁹ / l, lymphocytes 2.1 • 10 ⁹ / l, monocytes 0 , 2 • 10 ⁹ / l.

 Biochemical analysis of blood: total protein 90 g / l, fibrinogen 5 g / l, sialic acids 4.25 mmol / l.

Indicators of CL urine (Fig. 1):
1. July 12, 1999, at the time of the survey, Cn was 10.5 • 10 ⁶ quanta / min, A - 180.0 • 10 ⁵ quanta / min and S - 48.0 • 10 ⁵ quanta / min;
2. On July 20, 1999, during the course of treatment, Cn was equal to 9.5 • 10 ⁵ quanta / min, A - 183.0 • 10 ⁵ quanta / min, and S - 40.5 • 10 ⁵ quanta / min;
3. 02.08.99, at the discharge of Sp - 9.3 • 10 ⁵ quanta / min; A - 173.0 • 10 ⁵ quanta / min; S - 46.3 • 10 ⁵ quanta / min.

Indicators of CL blood (Fig. 2):
1. July 12, 1999, at the time of the survey, Cn was 4.5 • 10 ⁵ quanta / min, A - 144.0 • 10 ⁵ quanta / min and S - 23.0 • 10 ⁵ quanta / min;
2. On July 20, 1999, during the course of treatment, Cn was equal to 4.1 • 10 ⁵ quanta / min, A - 152.0 • 10 ⁵ quanta / min, and S - 26.0 • 10 ⁵ quanta / min;
3. 02.08.99, at discharge Sp - 3.4 • 10 ⁵ quanta / min; A - 155.0 • 10 ⁵ quanta / min; S - 22.0 • 10 ⁵ quanta / min.

 An in-depth clinical and laboratory examination revealed no abnormalities in chemiluminescence.

 In 15% of all patients, when examining CL, the blood was high, and urine, on the contrary, was low. During the complex treatment carried out at the sanatorium-preventorium, restoration of the CL and blood urine intensity was observed in 6.7% of cases.

 Example 2

 Fazlyeva A.N., 45 years old, works as an operator in OAO Ufaneftekhim. CL of urine on November 15, 1999, at the time of the examination, was 2 times lower than normal, and blood 2.5 times lower than normal. November 22, 1999 was admitted to a dispensary with a diagnosis of chronic lumbosacral radiculitis, neurocirculatory dystonia.

Complete blood count: hemoglobin 130 g / l, red blood cells 3.2 • 10 ¹² / l, CP 0.8, white blood cells 4.5 • 10 ⁹ / l, platelets 240 • 10 ⁹ / l, ESR 12 mm / h, stab 0.06 • 10 ⁹ / l, segmented 3.2 • 10 ⁹ / l, basophils 0.072 • 10 ⁹ / l, lymphocytes 2.2 • 10 ⁹ / l, monocytes 0.09 • 10 ⁹ / l.

 Blood enzyme activity: aspartate aminotransferase - 0.52 mmol / h • l, alanine aminotransferase - 0.86 mmol / h • l, creatine phosphokinase - 74 mmol of inorganic phosphorus / h • l.

Indicators of CL urine (Fig. 3):
1. On November 15, 1999, at the time of the examination, Cn was 4.5 • 10 ⁵ quanta / min,
A - 180.0 • 10 ⁵ quanta / min and S - 12.0 • 10 ⁵ quanta / min;
2. 22.11.99, during the treatment of Sp was equal to 17.7 • 10 ⁵ quanta / min,
A - 170.0 • 10 ⁵ quanta / min and S - 14.5 • 10 ⁵ quanta / min;
3. 08.12.99, at discharge Sp - 7.0 • 10 ⁵ quanta / min;
A - 185.0 • 10 ⁵ quanta / min; S - 18.2 • 10 ⁵ quanta / min.

Indicators of CL blood (Fig. 4):
1. November 15, 1999, at the time of the examination, Cn was 18.5 • 10 ⁵ quanta / min,
A - 203.0 • 10 ⁵ quanta / min and S - 86.0 • 10 ⁵ quanta / min;
2. 22.11.99, during the treatment of Sp was equal to 11.1 • 10 ⁵ quanta / min,
A - 244.0 • 10 ⁵ quanta / min and S - 62.0 • 10 ⁵ quanta / min;
3. 08.12.99, at discharge Sp - 6,2 • 10 ⁵ quanta / min,
A - 140.0 • 10 ⁵ quanta / min; S - 41.0 • 10 ⁵ quanta / min.

 During treatment with the use of stimulating therapy (extracts of eleutherococcus, ginseng, vitamin therapy, massage), restoration of blood and urine chemiluminescence was observed. Discharged (December 8, 1999) with improvement. Researches of CL of urine carried out repeatedly after 2-3 months of deviations from the norm did not show.

 A decrease to physiological values was noted in 69.4% of cases from this group.

 Example 3

 Tomina V.I., 52 years old, works as a freight forwarder at OAO Salavatnefteorgsintez. CL of urine and blood on 07/11/99, at the time of the examination was increased 2.5 times. Received July 17, 1999 with a diagnosis of hypertension 2 tbsp. , osteochondrosis of the cervicothoracic spine, chronic bronchitis.

Indicators of CL urine (Fig. 5):
1. July 11, 1999, at the time of the examination, Cn was 24.7 • 10 ⁵ quanta / min,
A - off scale. and S, 91.0 x 10 ⁵ quanta / min;
2. 17.07.99, during the treatment of Sp was equal to 14.1 • 10 ⁵ quanta / min,
A - 171.0 • 10 ⁵ quanta / min and S - 79.5 • 10 ⁵ quanta / min;
3. July 28, 1999, at the discharge of Sp - 9.2 • 10 ⁵ quanta / min;
A - 235.0 • 10 ⁵ quanta / min; S - 46.2 • 10 ⁵ quanta / min.

Indicators of CL blood (Fig. 6):
1. 07/11/99, at the time of the survey Sp was 22.4 • 10 ⁵ quanta / min,
A - off scale. and S, 76.0 x 10 ⁵ quanta / min;
2. 17.07.99, during the treatment of Sp was equal to 10.1 • 10 ⁵ quanta / min,
A - 177.0 • 10 ⁵ quanta / min and S - 59.0 • 10 ⁵ quanta / min;
3. 08.08.99, at discharge Sp - 3.3 • 10 ⁵ quanta / min;
A - 160.0 • 10 ⁵ quanta / min; S - 22.0 • 10 ⁵ quanta / min.

 In the course of treatment, sedatives, antioxidant therapy, including a complex of vitamins E and C, were used. At discharge normalization of blood and urine chemiluminescence was noted.

 Only in 6.9% during treatment and prophylactic measures there was no positive dynamics in the change in CL of blood and urine. Among this group of individuals (57 people), 11 were diagnosed for the first time with diseases such as diabetes mellitus, glomerulonephritis, pyelonephritis, cystitis, prostatitis.

 Example 4

 Ibragimova R.F., 38 years old, works as an operator in OAO Ufahimprom. ChL of urine on October 16, 1999, at the time of the examination was 3 times lower than normal, and blood 3.5 times lower than normal. 10.21.99, was admitted to a dispensary with a diagnosis of chronic pyelonephritis.

Complete blood count: hemoglobin 150 g / l, red blood cells 5.0 • 10 ¹² / l, CP 0.9, white blood cells 7.2 • 10 ⁹ / l, platelets 300 • 10 ⁹ / l, ESR 28 mm / h.

Protein fractions: globulins, α ₁ - 6.2 g / l, α ₂ - 7.8 g / l, β - 9.3 g / l.

 Residual nitrogen and its components in blood serum: residual nitrogen 18.2 mmol / L, urea 9.3 mmol / L, creatinine 0.2 mmol / L. Zimnitsky's test - a decrease in diuresis. The relative density of urine is 1030.

Indicators of CL urine (Fig. 7):
1. 16.10.99, at the time of the survey Sp was 3.3 • 10 ⁵ quanta / min,
A - 212.0 • 10 ⁵ quanta / min and S - 5.0 • 10 ⁵ quanta / min;
2. 21.10.99, during the treatment of Sp was equal to 5.0 • 10 ⁵ quanta / min,
A - 165.0 • 10 ⁵ quanta / min; and S - 10.5 • 10 ⁵ quanta / min;
3. 08.11.99, at discharge of Sp - 4.0 • 10 ⁵ quanta / min,
A - 200.0 • 10 ⁵ quanta / min; S - 5.0 • 10 ⁵ quanta / min.

Indicators of CL blood (Fig. 8):
1. 16.10.99, at the time of the survey Sp was 24.1 • 10 ⁵ quanta / min,
A - 277.0 • 10 ⁵ quanta / min; and S - 92.0 • 10 ⁵ quanta / min;
2. 21.10.99, during the treatment of Sp was equal to 10.6 • 10 ⁵ quanta / min,
A - 233.0 • 10 ⁵ quanta / min and S - 78.7 • 10 ⁵ quanta / min;
3. 08.11.99, at discharge Sp - 9.4 • 10 ⁵ quanta / min;
A - 268.0 • 10 ⁵ quanta / min; S - 51.3 • 10 ⁵ quanta / min.

 Example 5

 Kazantseva M.A., 42, has been an operator at ZAO Sterlitamak Petrochemical Plant. At the time of the examination, on 07.08.99, there was a sharp decrease in the intensity of CL in urine and an increase in the intensity of blood luminescence. Received August 15, 1999 with a diagnosis of chronic gastritis.

Indicators of CL urine (Fig. 9):
1. 07.10.99, at the time of the survey Sp was 5.0 • 10 ⁵ quanta / min,
A - 121.0 • 10 ⁵ quanta / min and S - 6.0 • 10 ⁵ quanta / min;
2. 15.10.99, during the treatment of Sp was equal to 7.0 • 10 ⁵ quanta / min,
A - 64.0 • 10 ⁵ quanta / min and S - 19.0 • 10 ⁵ quanta / min;
3. 28.11.99, at discharge of Sp - 5.0 • 10 ⁵ quanta / min;
A - 62.0 • 10 ⁵ quanta / min; S - 8.2 • 10 ⁵ quanta / min.

Indicators of CL blood (Fig. 10):
1. 07.08.99, at the time of the survey Sp was 10.9 • 10 ⁵ quanta / min,
A - 213.0 • 10 ⁵ quanta / min and S - 92.0 • 10 ⁵ quanta / min;
2. 15.08.99, during the treatment of Sp was equal to 8.1 • 10 ⁵ quanta / min,
A - 285.0 • 10 ⁵ quanta / min and S - 81.0 • 10 ⁵ quanta / min;
3. November 28, 1999, at discharge of Sp - 9.2 • 10 ⁵ quanta / min;
A - 264.0 • 10 ⁵ quanta / min; S - 57.0 • 10 ⁵ quanta / min.

 An in-depth examination of both Ibragimov R.F. and Kazantseva M.A. revealed diabetes mellitus.

Application of the proposed method will enable:
- evaluate the nonspecific reactions of the body's defense in the early stages of exposure, which is especially important when the concentration of "low intensity" and the combined effect of chemical pollutants;
- characterize the severity of adaptation mechanisms: from satisfactory (increase in spontaneous luminescence and CL light sum, normalization to baseline values, decrease in the light sum of urine luminescence, accompanied by a gradual increase in spontaneous luminescence of urine to a maximum) to disruption (a sharp increase in spontaneous luminescence and light sum of CL blood by a background of a decrease in spontaneous luminescence and light sum of CL urine);
- will reduce the incidence rate due to the timely implementation of high-quality preventive measures in people in contact with chemical pollutants of the working environment with low levels of spontaneous emission and light sum of urine CL and high levels of blood CL, which, combined with low costs for prediction, provides a decisive advantage of this method.

Claims (1)

A method for predicting a premorbid state when exposed to chemical pollutants of an industrial environment by examining urine, characterized in that the level of iron-induced chemiluminescence of urine and blood is recorded, the values of spontaneous luminescence and light sum of luminescence are calculated, and when the spontaneous luminescence in the blood is 2.0 • 10 ⁵ quanta / min and below and 7.0 • 10 ⁵ quanta / min and above, the luminosity of 10.0 • 10 ⁵ quanta / min and below and 40.0 • 10 ⁵ quanta / min and above, and in the urine spontaneous luminescence, equal to 6,0 • May ¹⁰ photons / m n and below and 12,0 • May ¹⁰ quanta / min or higher, the emission light sum equal to 16,0 • May ¹⁰ quanta / min and below and 70,0 • May ¹⁰ quanta / min and above, predict development premorbid state.

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