RU2329791C2 - Method of hemopoiesis stimulation associated with dogs' acute radiation sickness - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science. Method includes introduction of medicinal agent. Ceruloplasmin is used as specified medicinal agent. Prior to ceruloplasmin introduction for 7^th day after ionising irradiation, infra-red diode laser of power 25 W of wavelength 970 nm furnished with monofibre quartz light guide of diameter 0.4 mm, operating in pulse-periodic regime 100X50 is used for single osteoperforation procedure of scapula, iliac and ischial bones and breastbone, as well as epiphysis of humerus, radial, ulnar, femoral bones, shin bone and spellbone, and heel bone. Osteoperforation is carried out in 2-3 points by distance 1.0-1.5 cm from two sides during 2-3 seconds. Ceruplasmin is introduced drop-by-drop at rate 29-30 drops per minute once a day in daily dose 1.5 to 2.5 mg/kg of animal body weight. Course of ceruplasmin therapy is 7 procedures every 47-48 hours. It is established, that laser osteoperforation and ceruloplasmin intake provides intensive increase of erythrocyte, haemoglobin, leukocytes content in peripheral blood, as well as haemoglobin concentration in one erythrocyte. Medical efficiency is 100%.

EFFECT: development of effective method of hemopoiesis stimulation associated with radiation sickness in veterinary science.

2 tbl, 1 dwg

Description

The invention relates to the field of veterinary medicine and can be used to stimulate hematopoiesis in the treatment of acute radiation sickness (ARS) in farm and domestic animals.

Currently, various treatment regimens are used to treat ARS and stimulate bone marrow hematopoiesis in animals and humans.

A high-intensity laser is used to stimulate hematopoiesis in animals [6, 7, 8, 9].

A method has been developed to stimulate hematopoiesis in humans with ceruloplasmin [1, 10], used for anemia of various etiologies, including radiation therapy in the treatment of cancer patients.

In medical practice, treatment of ARS is aimed at normalizing blood formation organs (bone marrow transplantation, blood transfusion, administration of nucleic acid preparations, blood formation stimulants), fighting infection (antibiotics), preventing hemorrhages (vitamins), reducing intoxication (bloodletting, blood substitution), exposure to the nervous system [3, 4, 5].

However, the known method is not always effective, difficult to perform, as it involves complex surgical operations in case of bone marrow transplantation, or has undesirable side effects (rejection of bone marrow transplants).

There is a method of treatment and prevention of ARS in dogs, based on the use of indralin (preparation B-190) in combination with dimexide, pentovit, vicasol, antibiotics and the radioprotector indometophen [2].

The disadvantage of this method is its multicomponent nature and the absence of a pronounced stimulating effect on the function of bone marrow hematopoiesis.

The aim of the invention is to develop a method of stimulating bone marrow hematopoiesis in the treatment of ARS in animals.

This goal is achieved by the fact that in the claimed method, on the 7th day after irradiation, osteoperforations are applied once in the bones of the thoracic and pelvic extremities (scapula, ilium and sciatic bones), epiphyses of the tubular bones of the chest and pelvic limbs (humerus, radius, ulna, femur, tibia and tibia, calcaneus) on both sides and sternum for 2-3 seconds at 2-3 points for each pineal gland and limb belt bone at a distance of 1.0-1.5 cm from each other (drawing). They use an infrared diode laser in a pulse-periodic mode 100X50 with a wavelength of 970 nm, developing a power of 25 watts.

Energy was delivered transdermally, by contact, through a monofilament quartz fiber with a diameter of 0.4 mm. In this case, through perforations were formed in the area of the pineal glands in perpendicular planes.

Osteoperforation is carried out as follows. Puncture soft tissues to the bone with a needle with a diameter of 1.5 mm. A laser fiber is introduced into the needle cavity, laser osteoperforation is carried out. The time of osteoperforation (2-3 s) depends mainly on the breed, age and bone thickness in animals.

After osteoperforation, a course of treatment with ceruloplasmin in a daily dose of 1.5-2.5 mg / kg is carried out. The drug is administered dropwise at a rate of 29-30 drops per minute once a day, 7 injections with an interval of 47-48 hours.

Thus, in comparison with the prototype revealed the following distinctive features:

1. To stimulate hematopoiesis in ARS in animals, an infrared diode laser is used in a pulse-periodic mode 100X50 with a wavelength of 970 nm, developing a power of 25 watts.

2. Use a single osteoperforation of the bones of the thoracic and pelvic limbs (scapula, ilium and sciatic bones), epiphyses of the tubular bones of the thoracic and pelvic limbs (humerus, radial, ulnar, femoral, large and small tibia, calcaneal) from both sides and the sternum during 2-3 seconds at 2-3 points for each pineal gland and bone of the limb belt. The points are at a distance of 1.0-1.5 cm from each other.

3. After osteoperforation, a course of treatment with ceruloplasmin in a daily dose of 1.5-2.5 mg / kg is carried out. The drug is administered dropwise at a rate of 29-30 drops per minute once a day, 7 injections with an interval of 47-48 hours.

Patent studies have not revealed ways to stimulate hematopoiesis in the treatment of acute radiation sickness in animals using high-intensity radiation lasers and ceruloplasmin, characterized by the claimed combination of features, therefore, we can assume that this method meets the criterion of "novelty".

The use of a combination of essential features is also unknown, which indicates compliance with the criterion of "inventive step".

The inventive method can be carried out in any specialized veterinary institution, therefore, it meets the criterion of "industrial applicability".

The claimed method is illustrated by a description of its use and studies of the morphological composition of the blood.

Experimental studies were carried out on 12 outbred animals, selected according to the principle of analogues (age 2-3 years, weight - 13-14 kg, average fatness).

The animals were kept in individual cages under the conditions of vivarium UGAVM. Dogs were fed 3 times a day according to the norms established by the order of the Ministry of Health of the USSR No. 1179 of October 10, 1983. Water in the drinkers was constant. Dogs to maintain physiological functions receive a complete, balanced in basic elements feeding. The basis of their diet is feed of plant and animal origin. A typical diet for feeding dogs is shown in table 1 (gr. / Day for 1 animal).

Table 1. The ration of feeding animals by age groups (per animal) Animal species Product name milk groats potatoes cabbage carrot bread feed. yeast salt meat 2 cat. meat and bone meal barley oatmeal Adult dogs 200 one hundred one hundred 200 75 fifty 200 6 twenty 300 10

In the control and experimental groups (6 goals) to simulate acute radiation sickness, we used X-ray irradiation of animals on an X-ray-30 apparatus at a dose of 300 R. The irradiation was carried out at a voltage of 112 kV, a current of 40 mA, the total irradiation time was 36 with pulses of 2 from.

After osteoperforation in animals, a blood test was performed every 7 days until the 42nd day after the modeling of ARS. After irradiation in the control group, no treatment of animals was performed.

The dose was calculated by the formula:

D = U ² * I * t,

where D is the radiation dose, in mR;

U is the voltage in kV;

I is the current strength, in mA;

t is the total exposure time, in minutes.

In the control group (table 2), there was a sharp decrease in the leukocyte blood level, reaching a minimum value on the 21st day of observation - 1.71 ± 0.27 · 10 ⁹ L, which is 4.4 times lower than the initial value. Subsequently, there was a slow increase in the content of white blood cells, which on the 42nd day of observation was 2.90 ± 0.26 · 10 ⁹ / L, which nevertheless was 61.6% (2.6 times) lower than the background indicator.

The concentration of erythrocytes slightly decreased during the 28 days of observation and reached the minimum level of 5.23 ± 0.51 · 10 ¹² / l or 7.1% lower than the background value on the 28th day. Further, on the 42nd day of observation, an increase in the concentration of red blood cells was observed, which reached 5.29 ± 0.37 · 10 ¹² / L, which is 6.0% less than the background value.

The hemoglobin content in the blood and the concentration of hemoglobin in erythrocytes decreased during 42 days of observation and reached a minimum level of 110.26 ± 5.87 g / l and 221.46 ± 12.73 g / l, respectively, which is 4.5 and 8, 8% (P <0.05) below baseline.

In the experimental group of dogs (6 goals), on the 7th day after modeling radiation injury, an infrared diode laser was used in the treatment of ARS in a pulse-periodic mode, developing a power of 25 watts.

After osteoperforation, a course of treatment with ceruloplasmin in a daily dose of 1.5-2.5 mg / kg was carried out. The drug was administered dropwise at a rate of 29-30 drops per minute once a day, 7 injections with an interval of 47-48 hours.

After x-ray irradiation, the animals develop clinical signs of acute radiation sickness in the first 3 days, characterized by mild suppression, decreased food irritability, and in some cases vomiting. Then, within 5-7 days, the second phase of acute radiation sickness began and the dogs showed an improvement in their general condition, however, when conducting hematological studies, a sharp decrease in the level of leukocytes in the blood was revealed.

After laser osteoperforation in animals within 2 days there is a slight inhibition, a decrease in appetite. Body temperature at this time is held at the upper limit of the norm (39.0 ° C).

n=6)Table 2. Dynamics of hematological blood parameters of dogs of the experimental and control groups (

n = 6) Group BACKGROUND 3 7 fourteen 21 28 35 42 Red blood cells, 10 ¹² / L Control 5.63 ± 0.35 5.62 ± 0.49 5.31 ± 0.47 5.30 ± 0.40 5.26 ± 0.46 5.23 ± 0.51 5.28 ± 0.44 5.29 ± 0.37 Experienced 5.58 ± 0.32 5.59 ± 0.35 5.57 ± 0.34 6.34 ± 0.31 * 6.90 ± 0.25 * 8.07 ± 0.37 ** 7.13 ± 0.53 ** 7.22 ± 0.52 * White blood cells, 10 ⁹ L Control 7.56 ± 0.40 3.15 ± 0.30 2.21 ± 0.33 2.04 ± 0.25 1.71 ± 0.27 2.06 ± 0.29 2.31 ± 0.21 2.90 ± 0.26 Experienced 7.96 ± 0.64 3.26 ± 0.21 2.86 ± 0.34 1.94 ± 0.38 1.10 ± 0.37 2.31 ± 0.32 2.94 ± 0.36 4.32 ± 0.40 ** Hemoglobin, g / l Control 115.43 ± 4.12 115.81 ± 5.64 114.95 ± 4.67 113.64 ± 4.97 112.82 ± 5.84 112.18 ± 4.64 111.77 ± 4.31 110.26 ± 5.87 Experienced 114.92 ± 4.25 114.54 ± 5.77 115.26 ± 5.80 120.69 ± 6.26 125.49 ± 3.46 ** 131.84 ± 4.81 *** 127.49 ± 3.46 *** 126.05 ± 4.79 ** The concentration of hemoglobin in the red blood cell, g / l Control 242.90 ± 12.51 247.72 ± 15.32 243.55 ± 14.84 244.13 ± 20.92 232.31 ± 10.34 232.37 ± 12.38 228.14 ± 11.49 221.46 ± 12.73 Experienced 240.73 ± 18.38 235.14 ± 17.35 239.57 ± 12.76 243.43 ± 20.34 258.51 ± 10.19 * 267.32 ± 14.42 * 252.32 ± 13.12 * 259.21 ± 10.87 * significant changes compared to baseline: * - P <0.1 ** - P <0.05 *** - P <0.01

After 2 days, the condition of the animals normalizes, the body temperature decreases slightly (up to 38.5 ° C).

After x-ray irradiation in dogs there was a sharp decrease in the content of white blood cells in peripheral blood from 7.96 ± 0.64 · 10 ⁹ / L to 3.26 ± 0.21 · 10 ⁹ L on day 3 and 2.86 ± 0. 34 · 10 ⁹ l on the 7th day after irradiation. The content of red blood cells and hemoglobin remains at the same level (table 2).

On the 14th day of the experiment (7 days after laser osteoperforation and the start of treatment with ceruloplasmin) there was a further decrease in the blood leukocyte count, which reached a minimum value of 1.10 ± 0.37 · 10 ⁹ l on day 21. In the following days of observation, there was an increase in the content of leukocytes, the level of which reached 4.32 ± 0.40 · 10 ⁹ / L on the 42nd day of observation.

The concentration of erythrocytes in the blood increased from 14 to 28 days of observation and reached 8.07 ± 0.37 · 10 ¹² / L, which is 44.6% higher than the background value. Subsequently, the erythrocyte content decreased to the level of 7.22 ± 0.52 · 10 ¹² / l on the 42nd day of the experiment.

The hemoglobin content in the blood and the concentration of hemoglobin in the red blood cell correlate with the concentration of red blood cells. Starting from the 14th day of the experiment, there is an increase in the concentration of hemoglobin in the blood and the concentration of hemoglobin in the red blood cell, which reaches a maximum on the 28th day of observation (131.84 ± 4.81 g / l and 267.32 ± 14.42 g / l respectively), and then there is a decrease to the level of 126.05 ± 4.79 g / l and 259.21 ± 10.87 g / l, which is 9.7% and 7.7% higher than background values.

The clinical condition of the dogs of the control group continued to deteriorate for 35 days and 2 dogs (33.3%) died due to the development of septic processes.

In the experimental group of dogs, the clinical signs of acute radiation sickness were less pronounced and by the end of the observation period (42 days) all animals looked clinically healthy. Therapeutic efficacy was 100%.

Thus, the obtained results prove the high efficiency of osteoperforation of the pineal gland using a high-intensity infrared diode laser and ceruloplasmin to stimulate hematopoiesis in the treatment of acute radiation sickness.

Literature

1. Instructions for use of ceruloplasmin. Registration number 90/411/1. Approved by the Pharmacological Committee 10/19/1990

2. Kolesnichenko, I.S. Radiation prevention and treatment regimens for service dogs. / Kolesnichenko I.S., Mikhailov L.S., Boyarinov A.S., Grishin A.V. // Veterinary medicine, 2005; No. 12, - S. 52-54.

3. Kurshakov NA, Radiation sickness, in the book: A multivolume guide to internal diseases, v.10, M., 1963; Radiation medicine, 4th ed., M., 1968.

4. Krotkov F.G., Man and radiation, M., 1968.

5. Lindenbraten L.D., Medical Radiology, M., 1969.

6. Molokanov, V.A. Stimulation of hematopoiesis in animals by laser osteoperforation. / V.A. Molokanov, V.V. Strizhikov // Actual problems of biology and veterinary medicine of small pets: collection of books. scientific tr - Troitsk, 2005 .-- S.265-269.

7. Pashnin, D.A. The influence of high-intensity laser radiation on the dynamics of hematopoiesis and lipid peroxidation / D.A. Pashnin, V.V. Strizhikov, R.V. Mikhailenko // Young scientists - Russian agriculture. Materials of the II All-Russian Conference of Young Scientists (November 23-25, 2005): Sat. scientific tr - Moscow, 2005.

8. Pashnin, D.A. The effect of laser osteoperforation on the biochemical status of dogs. / D.A. Pashnin, V.V. Strizhikov, R.V. Mikhailenko // Promising areas of scientific research of young scientists. Materials of the IX scientific-practical conference dedicated to the 75th anniversary of the UGAVM (November 9-11, 2005): Sat. scientific tr - Troitsk, 2005 .-- C.113-114.

9. Strizhikov, V.V. The dynamics of hematopoiesis, the content of total protein and medium molecular peptides in blood serum on the background of laser osteoperforation. / V.V. Strizhikov, D.A. Pashnin, R.V. Mikhailenko // Promising areas of scientific research of young scientists. Materials of the IX scientific-practical conference dedicated to the 75th anniversary of the UGAVM (November 9-11, 2005): Sat. scientific tr - Troitsk, 2005 .-- S.142-143.

10. Pat. 2268745 Russian Federation, IPC A61K 38/43, A61K 33/34, A61K 33/14, A61N 5/10, A61P 7/06 Method for the prevention and (or) treatment of anemia in children (options). / Chardymova L.R., Parshikov V.V., Privalova L.P., Efremova L.M., Kraineva T.A .; applicant and copyright holder Nizhny Novgorod State Medical Academy. - 2003121205/14; declared 07/08/2003; publ. 01/10/2005, bull. No. 03.

Claims (1)

A method of stimulating hematopoiesis in the treatment of acute radiation sickness in dogs, comprising administering medicaments, characterized in that ceruloplasmin is used as a medicament, before which it is administered on the 7th day after ionizing irradiation with a 25 W infrared diode laser with a wavelength of 970 nm equipped with a monofilament quartz fiber with a diameter of 0.4 mm, in a pulse-periodic mode of 100 × 50 carry out a single osteoperforation of the scapula, iliac, sciatic bones and sternum, as well as epiphyses of the humerus, radius, ulna, femur, tibia, calcaneus and calcaneus at 2-3 points at a distance of 1.0-1.5 cm on both sides for 2-3 seconds, and ceruplasmin is administered dropwise at a speed of 29 -30 drops per minute once a day in a daily dose of 1.5 to 2.5 mg / kg of animal body weight, while the course of treatment with ceruplasmin is 7 treatments with an interval of 47-48 hours.