RU2219894C2 - Method of treating diabetes mellitus in children and adolescences - Google Patents

Abstract

FIELD: endocrinology. SUBSTANCE: after normalization of metabolism, biologically active points for introducing cellular suspension are found, which points are related to 4-6 damaged organs. Cellular suspension is prepared from mammal organs and is introduced in amounts 0.3-0.5 ml per injection, 2 to 5 injections being made for each damaged organ. EFFECT: reduced doses of administered insulin, prolonged remission terms, and stabilized diabetes mellitus course period. 1 tbl, 2 ex

Description

 The invention relates to medicine, namely to endocrinology and pediatrics.

 Insulin-dependent diabetes mellitus takes a significant place among chronic diseases of children and adolescents, reduces life expectancy and significantly increases the disability of young people. Diabetes mellitus in children and adolescents, especially in the pre- and puberty, is characterized by an unstable course with a tendency to ketoacidosis and hypoglycemic conditions, the inability to achieve stable metabolic compensation in most patients. This fact is explained by an increase in the level of contra-hormonal hormones, which leads to morphological and clinical manifestations, the progression of vascular changes, disability at a young age and early deaths.

 A known method of treating diabetes mellitus by administering prolonged and short acting insulins before a meal. Each patient has an individual dose distribution of insulin (M.A. Zhukovsky, 1995., E.P. Kasatkina, 1996., M.I. Balabolkin, 1998).

 However, in this way it is impossible to achieve stabilization of the disease, prevention of complications, it is impossible to provide an adequate level of life insulin levels.

 Closest to the proposed one is a method of treating diabetes mellitus by administering a drug based on cell suspension prepared from hematopoietic cells of the liver or spleen embryos taking into account the patient’s individual characteristics (Patent of Russia 2126260, 1999, M. Cl. A 61 K 31/28 )

 The use of this method is complicated by the problem of obtaining embryos and the risk of infectious diseases. In addition, the method does not have the ability to treat diabetes mellitus, and only affects the nature of the immune response. From the literature, we also have not received information on the effectiveness of this method for diabetes in children.

 The technical result of the proposed method is to ensure a stable course of diabetes, lower doses of insulin, lengthening the remission of the disease, ensuring regression of complications of the underlying disease.

 What is new in achieving a technical result is that after achieving normalization of metabolism, biologically active points of introduction of cell suspension are determined, which address 4-6 affected organs.

 Also new is the fact that cell suspension is injected subcutaneously in 0.3-0.5 ml and 2-5 injections for each affected organ.

 The points of introduction of cell suspension are determined according to the Voll method using a computer version (Handbook of representative points of electroacupuncture according to R. Voll. - Moscow: MC "System", 1991). At the same time, injection points are indicated that are addressed to the affected organs, which allows stabilizing the functions of this organ during treatment, which. in turn, ensures the stability of the course of diabetes and the regression of complications.

 The introduction of a mammalian cell suspension subcutaneously into biologically active points provides a large amount of exposure, which increases the effectiveness of treatment due to normalization of carbohydrate, lipid and protein metabolism, positive dynamics of physical and sexual development, as well as normalization of the psychoemotional state of a child and adolescent.

 The number of injected cell suspension and the number of injections per organ by the authors was determined empirically.

 Comparative analysis with the prototype shows that the inventive method is characterized in that after achieving metabolic normalization, the points of introduction of cell suspension are determined, addressing 4-6 affected organs, while the cell suspension is injected subcutaneously with 0.3-0.5 ml per injection and 2-5 injections per affected organ, which meets the criterion of "novelty."

 A new set of features ensures the stability of diabetes mellitus, lower doses of insulin, prolonged remission of the disease, positive dynamics of physical and sexual development, normalization of the psycho-emotional state, which meets the criterion of "industrial applicability".

 The method is carried out by the following steps.

 Preparing the patient for treatment is carried out within 7-10 days, the preparation consists in examining and achieving normalization of metabolism by traditional methods. The examination includes registration of clinical symptoms, instrumental methods of investigation (ultrasound of the abdominal cavity, kidney, thyroid gland, EEG and M-echo, ECG, ultrasound of the brain vessels and lower extremities, FLG, computer diagnostics by the Voll method, ophthalmoscopy). Laboratory studies include biochemical tests (sugar curve, daily glucosuria, glycosylated hemoglobin, C-peptide), lipid peroxidation and antioxidant protection of blood; immunological tests of the first level taking into account the absolute and relative content of immunocomplex cells (IR), receptor activity of lymphocytes in the E-ROCK, M-ROCK tests, theophylline (E-ROCK / senses, E-ROCK t / rez), phagocytic reactions (FAN, FI, FAK, NVT sp, NVT in), the content of immunoglobulins A, M, G according to Mancini. Compensation for diabetes includes the normalization of carbohydrate and lipid metabolism, treatment of complications of diabetes mellitus and concomitant diseases with antibiotics, angioprotectors, hepatoprotectors, membrane-stabilizing therapy, and vitamin therapy. The determination of the points of introduction of cell suspension is carried out according to the Voll method using a computer version. At the same time, biologically active points of administration, addressed to 4-6 affected organs, are noted, depending on the results of the examination. Marked dots are stained.

 To solve the question of the introduction of cell suspension using data from clinical, instrumental and laboratory studies.

For cell suspension, tissues of the pancreas, spleen, liver, adrenal glands, kidneys, myocardium, lung, and brain of a mammal, such as a newborn piglet, are used. When preparing cell suspension from the pancreas, remove the capsule of the organ, grind it into fragments of 2-3 ml, pour the cells with a collagenase solution, type 3 (Paneko NPP) at a concentration of 1 mg / 1 ml for 15 minutes at 37 ^o C and wipe through a sieve . The cell mixture is resuspended in a Hanks solution at 4 ^° C., and then centrifuged twice at 1000 rpm for 10 minutes. The precipitate is resuspended in Hanks solution. Tissue cells of other organs for xenograft are prepared according to the same scheme, changing the time of collagenase exposure to an organ, for example, liver, kidney, spleen cells are treated for 15 minutes, adrenal and myocardial cells - for 10 minutes, lung cells are treated with collagenase solution 25 minutes. The isolated cells are stored on ice until use. The vital activity of cell suspension is controlled using a test with a 1% trypan blue solution. The percentage of living cells is 95-99%.

 The introduction of the finished cell suspension is carried out in the operating room. The dose of injected cell suspension is individually selected from 0.3 to 0.5 ml per injection. The cell suspension is injected subcutaneously at the marked biologically active point, the total introduction of the cell suspension is 1.0-2.5 ml per organ. In treatment, cells from 4 to 6 organs are used. After transplantation, taking into account the patient’s condition, blood sugar is determined 4-5 times a day, daily glucosuria is determined. Strictly control the diet. According to the observations of the authors, hypoclycemia occurs on the first day in 100% of cases.

 Already on the first day after transplantation, blood sugar decreases (M + m = 12.1 ± 1.7 mmol / L before the introduction of cell suspension; 7.4 ± 0.1 mmol / L after 3-5 days) and remains within these limits within 8-12 months after transplantation of cell suspension. Accordingly, the dose of insulin administered decreased from 32 ± 2.5 to 18 ± 1.8 units per day. The dynamics of daily glucosuria varied from 84.8 g before the introduction of cell suspension to 146 g 3-5 days after the introduction of cell suspension. The level of C-peptide rises from 0.18 ± 0.06 to 0.52 ± 0.12 pg / ml one month after the introduction of cell suspension (0.8-4.0 pg / ml). Sugar stabilization at lower numbers than before treatment and at lower doses of insulin occurred within 30-45 days (Table 1).

 Upon admission, the true labile course of diabetes in children and adolescents is 100% of cases. Distal polyneuropathy was recorded in 62% of cases, manifested by convulsions, pain, parasthesia in the legs and feet. In all patients, these subjective sensations disappeared or became less intense during the first days after the introduction of cell suspension. The effect lasted 8-12 months. The stabilization of the course of labile forms of diabetes in children and adolescents was observed within 30-45 days after the introduction of cell suspension. After treatment, the proposed method in children and adolescents for 8-12 months, growth increased. Sexual signs corresponding to age were formed in 14 of 42 adolescents.

 The proposed method for the treatment of diabetes is illustrated by the following clinical examples.

Example 1. Kozlov Dima, 13 years old. The duration of diabetes mellitus is 4 months (course of diabetes with a tendency to frequent hypoglycemic conditions). The daily dose of insulin is 20 PIECES / day. Glycosylated hemoglobin 7.2%. In urine, sugar is 6 g / day. Blood sugar at admission 6.6-12.8-8.9-7.4 mmol / L. Height 147 cm, weight 35.5 kg. Sexual development of RoAx ₀ F ₀ V ₀ L ₀ . A cell suspension was prepared from the pancreas, liver, spleen, and adrenal glands of newborn piglets. In biologically active points, addressed to the affected organs, it was introduced: Langerhans islets - 0.5 ml 2 injections, liver 0.5 ml 3 injections, spleen 0.5 ml 2 injections, adrenal glands 0.3 0.3 injections.

3 months after treatment:
Height 152 cm, body weight 39 kg, blood sugar - 4.4-6.0-5.2-8.2 mmol / L. Glycosylated hemoglobin 6.2%. In the urine, aglucosuria, the daily dose of insulin is 8 units.

After 1 year, at 14, the second transplantation of cell suspension was performed. Upon receipt of the dose of insulin 20 units / day. Blood sugar 14.2-24.0-8.0-12.2 mmol / L. Glycosylated hemoglobin 7.9%, height 155 cm, weight 47.5 kg. Sexual development corresponds to Po ₁ Ax ₀ F ₀ V ₁ L ₁ age. A cell suspension was prepared from the pancreas, liver, spleen, kidney, adrenal gland of newborn piglets. In biologically active points, addressed to the affected organs, the following were introduced: Langerhans islets, 0.5 ml 2 injections, liver 0.5 ml 5 injections, spleen 0.5 ml 2 injections, adrenal glands 0.5 ml 2 injections, kidneys 0.5 ml 2 injections.

7 months after treatment, blood sugar 4.7-6.2-5.0-7.2 mmol / l, insulin dose 12 U / day, urinary aglucosuria, glycosylated hemoglobin 5.9%. C-peptide before treatment 0.2 pg / ml, one month after treatment 0.7 (normal 0.8-4.0 pg / ml). Height 161 cm, weight 54 kg. Sexual development P ₂ Ax ₁ F ₀ V ₂ L ₂ .

Example 2. Vakulenko Alexander, 7 years old. Diagnosis: diabetes mellitus insulin-dependent severe with a tendency to frequent hypoglycemic conditions. The duration of the disease is about 2 years. Upon receipt of glycemia - 6.9-7.3-10.2-5.2-13.2.2 mmol / L. Dose of insulin - 18 units / day. Glycosylated hemoglobin - 9.3%. Height 119 cm, body weight 20.5 kg. After normalizing the metabolism and determining the points of introduction addressing the affected organs, a cell suspension was prepared from the pancreas, liver, adrenal glands, and spleen of newborn piglets. Cell suspension was injected subcutaneously in the amount of: islets of Langerhans 0.5 ml 2 injections, liver 0.5 ml 3 injections, adrenal glands 0.3 ml 2 injections, spleen 0.5 ml 2 injections. On the third day after the introduction of cell suspension, blood sugar 5.8 = 6.5-6.2-10.2-7.8 mmol / L. Dose of insulin 15 units / day. 6 months after treatment, height 122 cm, weight 21 kg. 3 months after treatment, urine sugar is negative. The dose of insulin 0. Glycosylated hemoglobin 5.3%. After 6 months, height 127 cm, body weight 22 kg. Moved SARS with t 38 ^o C. Insulin 8 units / day, sugar in the urine 12 g / day. After 12 months, height 130 cm, body weight 22 kg. The daily dose of insulin is 14 PIECES. Glycosylated hemoglobin 7.3%.

 A total of 52 cell transplants were performed for children and adolescents aged 4.5 to 18 years with a disease duration of 4 months to 13 years.

Clinical indications for xenograft treatment:
- the labile course of diabetes;
- delayed physical and sexual development;
- the formation and progression of diabetic angiopathies;
- depletion of the effect of previous transplantation.

Thus, the proposed method for the treatment of diabetes provides:
- stabilization of diabetes from 8 months to 1 year or more;
- positive changes in physical and sexual development;
- normalization of neuropsychic function;
- the ability to achieve a more stable course of the disease against the background of the use of lower doses of insulin;
- prevention, regression of microangiopathies (nephropathy, retinopathy, encephalopathy, enteropathy, etc.).

Claims (1)

A method of treating diabetes mellitus in children and adolescents by introducing into the patient’s body a cellular suspension of mammalian organs, characterized in that, after achieving normalization of metabolism, biologically active points of introduction of cellular suspension are determined, addressing 4-6 affected organs, while the cell suspension is administered subcutaneously at 0 3-0.5 ml per injection and 2-5 injections per organ.

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