Method of non-pharmacological therapy of autistic spectrum disorders in children

Abstract

FIELD: neurology; psychiatry.SUBSTANCE: used as a non-pharmacological therapy for autism spectrum disorders in children. The method includes a set of procedures: interval hypoxic-hyperoxic training (HHT), hyperbaric oxygenation and ozone therapy. HHT consists of alternating inhalations of a hypoxic mixture with an oxygen content of 9 to 16 vol.% and a hyperoxic mixture with an oxygen content of 32 to 40 vol.% for a total duration of 45-50 minutes under the control of cardiovascular system parameters. Hyperbaric oxygenation is performed in a pressure chamber under a pressure of 1.3 atmospheres of 100% oxygen mixture, flow of 10 liters per minute, for 45 minutes using a mask. Ozone therapy is performed by rectal insufflation with an ozone-oxygen mixture with an ozone concentration of 85 μg / ml in a volume of 20–30 ml or by intravenous administration of a physiological solution enriched with ozone with an ozone concentration of 85 μg / ml at a rate of 200 ml per 100 ml of physiological solution. The procedures are carried out daily for 12 days.EFFECT: method is effective and has a positive impact on the patient’s condition.1 cl, 1 dwg, 3 ex

Description

[0001] The invention relates to the fields of psychiatry and mitochondrial medicine, namely, it concerns a method for non-pharmacological treatment of autism spectrum disorders in children with the complex use of hypoxic therapy, hyperbaric oxygenation and ozone therapy methods.

[0002] Autism spectrum disorders (ASD) are a group of disintegrative mental disorders characterized by a pronounced lack of social interaction and communication, limited interests and stereotypical behavior. According to the WHO, the global prevalence of ASD is currently 1 / 100 children, while in 2012 the median prevalence of ASD was approximately 1 / 160 children. [Zeidan J. et al. Global prevalence of autism: A systematic review update / / Autism research. - 2022. - V. 15. - No. 5. - P. 778-790.] Thus, the epidemiological picture of the prevalence of ASD continues to rapidly worsen.

[0003] The heterogeneity of the clinical picture and identified biological pathologies in patients causes uncertainty in the general etiopathogenetic concept of ASD. Leading studies on the pathogenetic mechanisms of ASD development determine the leading role of immunological and neurotransmitter disorders in patients. The concept of neuroinflammation unites a chain of neuro-immuno-biochemical disturbances leading to the development of ASD. One of the fundamental mechanisms combining inflammatory and pathobiochemical processes is mitochondrial dysfunction. [Rossignol DA, Frye R.E. Evidence linking oxidative stress, mitochondrial dysfunction, and inflammation in the brain of individuals with autism / / Frontiers in physiology. - 2014. - V. 5. - P. 150.]

[0004] Primary mitochondrial pathologies associated with genetic defects in mitochondrial protein synthesis (MELAS syndrome, Kearns-Sayre syndrome, Leigh syndrome, etc.) are known to be accompanied by mental disorders, along with peripheral and systemic manifestations. The most common are cognitive impairment, seizures, motor and speech delays, and, less commonly, psychotic episodes. Mental disorders combined with central and peripheral motor impairments are referred to as mitochondrial encephalopathy.

[0005] Mitochondrial dysfunction is also considered as a component of the pathophysiology of a number of mental and neurodegenerative diseases, in particular schizophrenia, major depressive disorder, bipolar disorder, ASD, Alzheimer's disease, Parkinson's disease, multiple sclerosis and others. [Ni P., Ma Y., Chung S. Mitochondrial dysfunction in psychiatric disorders / / Schizophrenia Research. - 2022., Bustamante-Barrientos FA et al. Mitochondrial dysfunction in neurodegenerative disorders: Potential therapeutic application of mitochondrial transfer to central nervous system-residing cells / / Journal of Translational Medicine. - 2023. - V. 21. - No. 1. - P. 613.]

[0006] According to the meta-analysis [Rossignol DA, Frye RE Mitochondrial dysfunction in autism spectrum disorders: a systematic review and meta-analysis / / Molecular psychiatry. - 2012. - V. 17. - №. 3. - P. 290-314.] mitochondrial dysfunction is widespread among patients with ASD and is secondary (not genetically determined) in nature. Among patients with early childhood autism, mitochondrial dysfunction was noted in more than 87% of cases, and about a third of patients had signs of decompensated systemic mitochondrial dysfunction. [Sukhorukoy V.S., Kireeva I.P. Possibilities of energy-tropic therapy in the complex treatment of early childhood autism / / Effective pharmacotherapy. - 2013. - №.50. - P. 4-10.]

[0007] Mitochondrial dysfunction is accompanied by a decrease in the energy-producing function of the cell, the development of oxidative stress, and changes in the regulation of cellular apoptosis.

[0008] The central nervous system is highly energy-dependent. CNS tissues contain a large number of mitochondria, making them extremely vulnerable to energy metabolism deficits and disruptions in redox homeostasis.

[0009] Adequate energy supply to neurons is critical for maintaining membrane potentials and transmitting nerve impulses.

[0010] Oxidative stress leads to disruption of the functional and structural integrity of neurons, as well as dysregulation of ROS signaling mechanisms.

[0011] A decrease in the activity of a number of enzymes (adenylate cyclase, Na+ / K+-ATPase, Ca2+-ATPase) of cellular membranes under conditions of energy deficiency during oxidative stress leads to membrane depolarization, disruption of water-electrolyte metabolism and pH, resulting in the destruction of the phospholipid layer of cellular membranes, and damage / shortening of mtDNA. In this case, a vicious circle is observed: oxidative stress leads to mtDNA mutations, and the accumulating mutated mtDNAs that have escaped autophagy favor the further development of oxidative stress [Velskikh E.S., Zvyagina V.I., Uryasyev O.M. Modern concepts of the pathogenesis and approaches to the correction of mitochondrial dysfunction / / Science of the Young - Eruditio Juvenium. - 2016. - No. 1. - P. 104-112.]. Shortened copies of mtDNA not only successfully compete with full-fledged ones for limited intracellular resources, but also reproduce faster than them.This explains the phenomenon of “clonal expansion of mutated mtDNA”, accompanied by the accumulation of aberrant mitochondria.

[0012] Under conditions of intracellular acidification, disruptions in the biochemical pathways of neurotransmitter metabolism also occur, leading not only to neurotransmitter imbalance but also to the synthesis of toxic metabolites. For example, oxidative stress is known to increase glutamate excitotoxicity, one of the proposed mechanisms for the development of ASD symptoms.

[0013] Violation of the structural integrity of cells, increased synthesis of aggressive and toxic metabolites, excitotoxicity, and disruption of calcium homeostasis lead to the launch of an apoptotic program and shift the balance of neurogenesis and synaptic plasticity processes towards neurodegeneration.

[0014] Thus, insufficient energy production, oxidative stress, metabolic disorders, dysregulation of synaptic transmission, apoptosis are factors that contribute to the disruption of the central nervous system, which are especially clearly reflected in the formation of the developing brain.

[0015] Mitochondrial dysfunction and accompanying inflammation are generalized in nature and cause systemic manifestations of ASD: non-specific constitutional symptoms (fatigue, muscle hypotonia, etc.), gastrointestinal disorders, endocrine disorders, etc.

[0016] Today, despite multiple studies confirming neuroimmunological and biochemical disorders in patients with ASD, ASD therapy is based on psychological and pedagogical correction. Drug therapy is indicated for significant manifestations of behavioral deviations or in the presence of comorbid conditions and is based on the use of neuroleptics, antidepressants and nootropic drugs. The effectiveness of the groups of drugs used by patients has been proven only in isolated studies. [Bastin J. Exposure to resveratrol triggers pharmacological correction of fatty acid utilization in human fatty acid oxidation-deficient fibroblasts / J. Bastin, A. Lopes-Costa, F. Djouadi / / Hum Mol Genet. - 2011. - Vol. 20, No. 10. - P. 2048-2057.; Dubinina EE Products of oxygen metabolism in the functional activity of cells (life and death, creation and destruction). Physiological and clinical-biochemical aspects / E.E. Dubinina. - St. Petersburg: Publishing house "Medical Press", 2006.- 400 pp., Ajith T.A. Mitochondria-targeted agents: Future perspectives of mitochondrial pharmaceutics in cardiovascular diseases / TA Ajith, TG Jayakumar / / World J Cardiol. - 2014. - Vol. 6, No. 10. - P. 1091-1099.

[0017] Given the significant pathogenetic role of mitochondrial dysfunction in the development of ASD, its correction is a necessary component of therapy. Correction of mitochondrial dysfunction is based on replenishing cellular energy deficits through the use of energy-stimulating therapy, as well as antioxidants, antihypoxants, vitamins, and vitamin-like supplements.

[0018] There is evidence of the effectiveness of metabolic therapy for the correction of mitochondrial dysfunction in patients with ASD. Thus, the use of complex energotropic therapy (L-carnitine, coenzyme Q10, acetylaminosuccinic acid) in children with early childhood autism contributed to the improvement of symptoms: a decrease in the severity of stereotypical movements, a decrease in hyperactivity, improvement of cognitive functions, an increase in concentration, an increase in the volume of active speech. [Krapivkin A.I., Sukhorukov V.S., Kireeva I.P. Possibilities of energotropic therapy in the complex treatment of early childhood autism / / Effective pharmacotherapy. - 2013. - No. 50. - P. 4-10.] Also, the use of metabolic therapy contributed to the improvement of laboratory parameters of mitochondrial function, and the clinically best results were obtained in the group of children with a decompensated form of systemic mitochondrial dysfunction [Krapivkin A.I., Sukhorukov V.S., Kireeva I.P.Potential of energy-tropic therapy in the complex treatment of early childhood autism / / Effective pharmacotherapy. - 2013. - No. 50. - P. 4-10.] Despite the positive results of treating patients with ASD using energy-tropic therapy, this method is only a supportive measure compensating for mitochondrial dysfunction, but does not allow for sustainable normalization of the mitochondrial pool.

[0019] Correction of the patient's condition should be systemic and include, along with the treatment of mitochondrial dysfunction, also anti-inflammatory, complex metabolic and hormonal therapy.

[0020] Non-pharmaceutical physiotherapy methods are equally effective. Well-researched and effective among these methods are hypoxic therapy, hyperbaric oxygenation (HBO), and ozone therapy. These methods are used to increase the body's nonspecific resistance, treat mitochondrial dysfunction, and treat infectious and inflammatory diseases. Thus, hypoxic therapy, HBO, and ozone therapy have many applications in the treatment of patients with ASD.

[0021] Hypoxytherapy is based on changing the composition of the inhaled gas mixture. The duration of hypoxic exposure is determined by the average value of the interval from the maximum to the minimum of the hypoxic cycle, after which the hypoxic exposure ceases until the next maximum. Technically, interval hypoxic therapy is carried out in the form of inhalations of a gas mixture with varying oxygen concentrations. The device is equipped with sensors that allow for the simultaneous assessment of parameters such as hemoglobin saturation with oxygen, heart rate, blood pressure, and ECG. There are 4 main training levels: A ("athletes"), B ("healthy people"), C ("sedentary people"), and D ("elderly, weakened people"). A single training session includes 4-6 cycles of breathing with hypo- and hyperoxic mixtures, the total time of breathing with the hypoxic mixture is 20-30 minutes.The regimen is selected based on the patient's individual tolerance to hypoxia following a hypoxic test or a trial training session at level D1. The parameters of the phases, frequency, and volume of respiration are adjusted to achieve increased tolerance to hypoxic stress while reducing the heart rate. Only then can hypoxic therapy be considered effective. As the body adapts to hypoxia, the regimens are adjusted toward those with a longer hypoxic phase and lower oxygen content in the gas mixture.

[0022] Hypoxytherapy increases the compensatory abilities of the body, corrects the functional state of patients. A number of literary sources describe in detail the relationship between changes in oxidative modes and correction of mitochondrial dysfunction. [Agadzhanyan N.A., Efimov A.I. Functions of the body under conditions of hypoxia and hypercapnia / / M., Medicine, 1986, 269 p.; Kolchinskaya A.Z. Mechanisms of action of IHT / / Hypoxia Medical J., 1993, No. 1, pp. 5-8; Velskikh E.S., Zvyagina V.I., Uryasyev O.M. Modern concepts of the pathogenesis and approaches to the correction of mitochondrial dysfunction / / Science of the Young - Eruditio Juvenium. - 2016. - No. 1. - P. 104-112., Tsyganova T.N., Prokopov A.F. Scientific basis for using the hypo-hyperoxytherapy method in the practice of mitochondrial medicine / / Physiotherapist. - 2016. - No. 3. - P. 15-22.The mechanism underlying the effectiveness of hypoxic therapy in the treatment of autism spectrum disorders is primarily associated with the correction of mitochondrial dysfunction and regulation of membrane permeability. Normalization of mitochondrial function is explained by the selective elimination (mitophagy) of damaged mtDNA [Prokopov, AF (2012). Intermittent Hypoxia and Health: From Evolutionary Aspects to Mitochondria Rejuvenation. In: Xi, L., Serebrovskaya, T. (eds) Intermittent Hypoxia and Human Diseases. Springer, London, https: / / doi.org / 10.1007 / 978-l-4471-2906-6_21], as well as increased cellular stability due to a decrease in the activity of lipid peroxidation in membranes. Simultaneously with the restructuring of the oxidative phosphorylation chain, the process of activation of anaerobic glycolysis occurs, which contributes to the energy supply of the body, and also increases the overall endurance to the effects of damaging factors [Berezovsky V.A., Volobuev M.I.Training to oxygen deficiency as a method for optimizing mental performance. Proc. VII international. symposium. "Ecological and physiological problems of adaptation". Moscow, 1998.

[0023] Both in domestic and foreign literature, there is a description of the practical application of hypoxic training to increase the plasticity and mobility of nervous processes, and as a consequence - to improve mental performance, [Kolesnikov A.N., Dubovaya A.V., B2 Naumenko Yu. PROSPECTS FOR THE USE OF NORMOBARIC HYPOXYTHERAPY IN CHILDREN (LITERATURE REVIEW) / / Bulletin of emergency and reconstructive surgery. - 2019. - Vol. 4. - No. 1. - P. 77-80.; Shilova O.V. Experience of using intermittent normobaric hypoxic therapy in the treatment of neurotic disorders / / Medical news. - 2005. - No. 11. - P. 104-106.], resistance to psycho-emotional stress / depression, increased productivity of mental work and concentration, academic performance, and a decrease in the number of errors [Adamyan N.Yu., Karapetyan M.A. Hypoxytherapy - treatment of oxygen deficiency / / - 2018. - V. 58. - No. 3. - P. 42-49]. improvement of short-term memory and concentration [Wang H, Shi X, Schenck H, et al. Intermittent Hypoxia Training for Treating Mild Cognitive Impairment: A Pilot Study. American Journal of Alzheimer's Disease & Other Dementias®. 2020;35. doi:10.1177 / 1533317519896725].

[0024] Hyperbaric oxygenation is a treatment method in which the patient is exposed to oxygen at elevated pressure in a special hyperbaric chamber. [Levina O.A. et al. Hyperbaric oxygenation in acute diseases and brain damage. New possibilities. New solutions / / Neurosurgery. - 2014. - No. 4. - P. 9-15.]

[0025] Hyperbaric oxygenation is based on increasing the partial pressure of oxygen (pO2) in the body's fluids (plasma, lymph, tissue fluid). Technically, there are several operating modes of the pressure chamber - a mode with an isopressure of 1.8-2 atm. and a mode up to 1.5 atm. [Baidin S.A., Gramenitsky A.B., Rubinchik B.A. Handbook of Hyperbaric Medicine / / M.: Medicine. - 2008. - V. 560] The HBO mode with a lower working pressure (up to 1.5 atm.) activates aerobic metabolism - this leads to a corresponding increase in oxygen capacity and is accompanied by an increase in oxygen diffusion into hypoxic areas of tissue, which makes it possible to fully satisfy the tissue's need for oxygen. In general, the therapeutic effect of HBO is due to the ability to compensate for oxygen deficiency (and, as a consequence, energy deficiency).

[0026] The use of hyperbaric oxygen also has immunomodulatory properties.

[0027] Hyperbaric oxygenation is an effective method of correcting neurological disorders, the functional state of the human body, maintaining, improving and restoring its performance, and this is evidenced by the results of both domestic and foreign researchers [Karimova L.A. The method of hyperbaric oxygenation in complex treatment / / Educational Bulletin "Consciousness". - 2012. - V. 14. - No. 1. - P. 12-13.]

[0028] Medical ozone is a universal stimulant. When applied locally (rectally), ozonated solutions have a pronounced anti-inflammatory and regenerative effect, but the leading effect remains immunomodulatory (phagocytosis activation is pronounced) [Bolgaev A.B. et al. ON THE QUESTION OF THE USE OF OZONE THERAPY IN MEDICINE / / International Bulletin of Medical Sciences and Clinical Research. - 2023. - V. 3. - No. 7. - P. 7-9.] Ozone activates a number of biochemical processes that result in the release of ATP, and is also an antihypoxant - it reduces the effects of tissue hypoxia and normalizes metabolic processes within the latter. When administered intravenously, it exhibits a hypocoagulant effect - it increases anticoagulant and fibrinolytic activity, reduces platelet aggregation (the ozone concentration is about 1 mg / l).The methods of application are not limited to intravenous; there are also intramuscular, subcutaneous, intravenous and rectal injections of ozonized solution, exposure to ozonized oil, ozonized water. Before intravenous administration, the physiological solution is ozonized [Minenkov A.A. et al. Basic principles and tactics of ozone therapy: A manual for doctors / / Moscow. - 2001. - P. 40.] For pediatric patients, the method of rectal insufflation of a gaseous ozone-oxygen mixture, carried out using a Janet syringe, is often used.

[0029] Patented sources of information contain information on the use of ozone for the treatment of patients with infectious diseases, for example, with herpes virus infection (RU 2178699, 04.04.2001). Ozone is widely used in various fields of medicine - dentistry, gastroenterology, dermatology, cardiology, neurology [Ozone therapy in neurology / A.V. Gustov, S.A. Kotov, K.N. Kontorshchikova, Yu.P. Potekhina. - Nizhny Novgorod: Information center for cooperation "Litera", 1999. - 243 p. - ISBN 5-900915-31-X. - EDN YLRXRZ.] and has proven itself as a reliable method of treatment with the correct selection of dosages and route of administration [Kulikov A.G. Ozone therapy is an integral part of physiotherapy / / Physiotherapy, balneology and rehabilitation. - 2005. - No. 4. - P. 3-7.] In neurology, the use of ozone therapy is widespread in relation to patients with chronic cerebral ischemia, cerebrovascular diseases, encephalomyelitis, dysmetabolic polyneuropathies [Belyakov K.M. Dysmetabolic polyneuropathies: clinical and electroneuromyographic diagnostic criteria, pathogenesis, new methods of restorative therapy / / Doctoral dissertation. - 2009.: Cherevaschenko LA et al. Ozone therapy and iodine-bromine baths in the correction of cognitive impairment in patients with chronic cerebral ischemia / / Spa medicine. - 2016. - No. 4. - P. 36-41. Davronov LO, Niyozov Sh.T., Dzhurabekova AT Treatment of encephalomyelitis and myelitis in children with ozone therapy / / Editor-in-chief: Sukiasyan AA, Cand. of Environmental Sciences, Senior Lecturer. - 2015. - P. 190.].

[0030] Thus, each of the described methods finds its application in various fields of medicine as an independent therapeutic tool.

[0031] Experience with the use of HBO in the treatment of ASD is known. During treatment with HBO in patients with ASD, an improvement in cognitive functions and social interaction was noted [Fischer I, Barak B. Molecular and Therapeutic Aspects of Hyperbaric Oxygen Therapy in Neurological Conditions. Biomolecules. 2020; 10(9): 1247. https: / / doi.org / 10.3390 / biom10091247]. Despite the fact that the use of HBO in ASD leads to an improvement in physiological and neuropsychiatric parameters, its effectiveness has been demonstrated only in a few studies. [Sakulchit T., Ladish C., Goldman RD Hyperbaric oxygen therapy for children with autism spectrum disorder / / Canadian Family Physician. - 2017. - V. 63. - No. 6. - P. 446-448.] Researchers conclude that HBO is safe and potentially effective, but not a comprehensive treatment method for children with ASD. [Sakulchit T., Ladish S., Goldman RD Hyperbaric oxygen therapy for children with autism spectrum disorder / / Canadian Family Physician. - 2017. - T. 63.- No. 6. - P. 446-448., Rossignol DA et al. Hyperbaric oxygen treatment in autism spectrum disorders / / Medical Gas Research. - 2012. - V. 2. - No. 1. - P. 1-13., Fischer I, Barak B. Molecular and Therapeutic Aspects of Hyperbaric Oxygen Therapy in Neurological Conditions. Biomolecules. 2020; 10(9): 1247. https: / / doi.org / 10.3390 / biom10091247] Probably, the limited effectiveness of HBO in ASD is due to the fact that the method was used as monotherapy. Taking into account the vastness of pathobiochemical processes involved in the development of ASD, the need for a combined use of various therapeutic tools to achieve a sustainable positive treatment outcome becomes obvious.

[0032] A study of patent literature revealed no cases of the combined use of hypoxic therapy, hyperbaric oxygenation and ozone therapy for the treatment of ASD.

[0033] The objective of the present invention is to introduce into practice a reliable, predictable, safe non-pharmacological method for correcting ASD.

[0034] The technical result of the invention is the positive dynamics of treatment of patients with ASD with improvement of the clinical picture and laboratory parameters.

[0035] The technical result is achieved through the use of a developed integrated method combining hypoxic therapy, HBO, and ozone therapy. Adequate and effective dosages, administration routes, as well as the duration and frequency of treatment courses, have been selected. This integrated approach allows for the renewal of the mitochondrial pool, thereby increasing the effectiveness of oxidative processes and eliminating energy deficiency. The anti-inflammatory, immunomodulatory, and disinfectant properties of the methods comprising this integrated therapy are additional factors enhancing its effectiveness. Stimulation of the neuroendocrine system, which occurs under the influence of various oxygenation regimens, contributes to an increase in the body's overall nonspecific resistance. Thus, the integrated use of hypoxic therapy, HBO, and ozone therapy addresses the key pathophysiological mechanisms of ASD.

[0036] The treatment course consists of hypoxic therapy, hyperbaric oxygenation, and ozone therapy procedures daily for 12 days. The hypoxic therapy method used is interval hypoxic-hyperoxic training, consisting of alternating inhalations of a hypoxic (oxygen content from 9 to 16 vol.%) and hyperoxic (oxygen content from 32 to 40 vol.%) mixture in various modes while monitoring cardiovascular parameters (such as heart rate and saturation). The choice of mode was based on physiological parameters and the level of hypoxia resistance. Gradually, the modes were changed towards increasing the duration of hypoxic intervals during hypoxic training. Hyperbaric oxygenation was performed in a pressure chamber under a pressure of 1.3 atmospheres of 100% oxygen mixture at a flow rate of 10 liters per minute using a mask. The procedure took 45 minutes, depending on the patient's condition and clinical situation, the procedure could last from 15 to 60 minutes.Ozone therapy for pediatric patients was administered by rectal insufflation of 20-30 ml of an ozone-oxygen mixture with an ozone concentration of 85 μg / ml through a special plastic tip using a Janet syringe. Alternatively, intravenous ozonated saline with an ozone concentration of 85 μg / ml was administered at a rate of 200 ml per 100 ml of saline solution for a total volume of 200-300 ml.

[0037] The effectiveness of the course of therapy is assessed by the dynamics of biochemical indicators of mitochondrial function (lactate and pyruvic acid in urine, organic acids in urine), by the dynamics of clinical symptoms and by resistance to hypoxia, assessed by changes in training regimens.

[0038] Figure 1 shows the protocols for 9 days of hypoxic-hyperoxic interval training. During this time, the training regimen was changed from level C2 to level B3, at which the patient maintained oxygen saturation. The hypoxic training index (HTi), which quantitatively reflects the total hypoxic load, increased from 49 in the first session to 153 in the ninth, reflecting the patient's increased tolerance to hypoxia.

[0039] The integrated method was tested on pediatric patients with ASD. The diagnosis of ASD was made based on ICD-11 criteria by a psychiatrist after a consultation. The severity of the patients' condition corresponded to the moderate to severe degree of autistic traits on the CARS scale. All patients were prescribed a 12-day course of integrated therapy. After the course, the patients demonstrated positive dynamics of laboratory parameters of mitochondrial function (initially elevated levels of lactate and pyruvic acid in urine decreased by 50% or more, normalization of other organic acids in urine reflecting metabolic processes), positive dynamics of the clinical picture (improvement of cognitive functions, social interaction, speech production, reduction of stereotypical movements, reduction in the severity or complete absence of undesirable behavior, stabilization of emotional lability, etc.).), increasing resistance to hypoxia (stable maintenance of saturation levels with increasing hypoxic load).

[0040] Thus, the proposed course of complex therapy is an effective way to treat patients with autism spectrum disorders.

[0041] The method is illustrated by the following clinical examples.

[0042] Example 1.

[0043] 4-year-old boy. Diagnosis: Autism spectrum disorder. Born at 39 weeks, with umbilical cord entanglement, Apgar score of 8 / 9. Weight 3600 g. Breastfed until 1 month, then mixed fed until 2.5 months, then bottle fed. Early development according to age. Age-appropriate immunizations. Until 1 year, repeated words and syllables, followed requests. At 1 year, after an acute respiratory viral infection (febrile temperature for 3 days, catarrhal symptoms for 7 days), speech skills regressed. From 1 year and 3 months, he stopped repeating words. He began to walk on his toes. Eye contact was lost. He stopped communicating with other children. He did not point before.

[0044] Received nootropic therapy: Tenoten, Semax, Gliatilin, Nootropil, Anvifen. Bioacoustic correction procedures were performed twice. As of October 2022: does not understand everything that is said to him. Becomes angry when asked. Does not speak (speaks his own language). Inordinate food intake. Does not ask to go to the toilet: is placed on a potty for urination, defecating in his pants. Constipation lasting up to 3 days. Sleep is not disturbed. Expressed food selectivity.

[0045] Mild periorbital cyanosis. Nails are peeling and brittle. Behavior includes agitation and mood swings. Pronounced stimuli and stereotypies. Decreased performance. Rapid fatigue. Previously suffered from atopic dermatitis. EEG with sleep (2022) - normal.

[0046] Test results from October 2022: S100 0.088 μg / L, Neuron-specific enolase 26.7 μg / L, ASLO normal, Eosinophilic cationic protein normal, Calprotectin normal, Herpes simplex virus type 6 not detected in oropharyngeal scraping. Gut microbiota - increased Staph. epidermidis, Blautia coccoides, Eggerthella lenta, Kingella. Acylcarnitine profile normal.

[0047] Organic acids in urine: lactate 67 (hereinafter, the measurement is in mmol / mol creatinine), pyruvic acid 54. Fumaric 2.7. 3-hydroxybutyric 37. Malonic 1.9. Glycolic 32. Quinolinic / xanthurenic acid ratio 18.

[0048] The patient underwent a 12-day course of daily procedures: interval hypoxic-hyperoxytherapy, hyperbaric oxygenation, and rectal ozone therapy.

[0049] Intermittent hypoxic-hyperoxytherapy:

[0050] The mode selection was based on the level of immersion in hypoxia and heart rate indicators.

[0051] Interval hypoxia-hyperoxytherapy therapy started with the "D4" mode: 2 minutes of breathing with a mixture containing 14% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen, 3 minutes of breathing with a mixture containing 14% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen, 4 minutes of breathing with a mixture containing 14% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen, 3 minutes of breathing with a mixture containing 13% oxygen, 5 minutes of breathing with a mixture containing 30% oxygen, 4 minutes of breathing with a mixture containing 13% oxygen, 5 minutes of breathing with a mixture containing 30% oxygen, 4 minutes of breathing with a mixture containing 13% oxygen, 5 minutes of breathing with a mixture containing 30% oxygen. The duration of the procedure is 47 minutes.

[0052] Over the course of 12 days, due to the body’s adaptation to hypoxia, the regimes changed towards regimes with an increasingly longer duration of the hypoxia stage with a lower oxygen content in the gas mixture.

[0053] By day 12, the "C3" mode was achieved: 4 minutes of breathing a mixture with an oxygen content of 14%, 4 minutes of breathing a mixture with an oxygen content of 30%, 5 minutes of breathing a mixture with an oxygen content of 14%, 4 minutes of breathing a mixture with an oxygen content of 30%, 5 minutes of breathing a mixture with an oxygen content of 14%, 4 minutes of breathing a mixture with an oxygen content of 30%, 5 minutes of breathing a mixture with an oxygen content of 14%, 3 minutes of breathing a mixture with an oxygen content of 30%, 5 minutes of breathing a mixture with an oxygen content of 14%, 3 minutes of breathing a mixture with an oxygen content of 30%, 5 minutes of breathing a mixture with an oxygen content of 13%, 3 minutes of breathing a mixture with an oxygen content of 30%. The duration of the procedure is 50 minutes.

[0054] Hyperbaric oxygenation was carried out in a pressure chamber with a pressure of 1.3 atmospheres, using a 100% oxygen mixture, a flow of 10 liters per minute using a mask.

[0055] Ozone therapy was carried out rectally in a volume of 30 ml of one-time ozone administration.

[0056] During therapy, developmental progress was noted: the appearance of individual syllables in speech, improved understanding of spoken language, and a decrease in the number of stimuli. A decrease in the severity of periorbital cyanosis was observed.

[0057] Example 2.

[0058] Patient, 4.5 years old. Diagnosis: Autism spectrum disorder. Born on time, natural delivery. Discharged on time. Early motor development is age-appropriate. Age-appropriate vaccinated, tolerated vaccinations well. Sickness is rare. No allergic processes were observed. Until the age of 3, development was age-appropriate. By the age of 3, he spoke individual words, but not phrases. There was no expressive speech. He poorly understood spoken language. He did not play with other children. He used pointing gestures. There was eye contact. There were no obvious pronounced stereotypies. Since September 2022, he has been studying using the ABA method. As of January 2023 (4.5 years old), the child has a large vocabulary (points to cards, can pronounce), but there is no addressed phrasal speech. Frequent echolalia. He understands spoken language and partially follows requests. He interacts actively with his family and has a strong need for physical contact. He doesn't play with children outside or in kindergarten. He doesn't approach them himself. If approached, he usually ignores them. He enjoys listening to classical music.He is demanding of protocol adherence: he walks a familiar route and requires objects to be arranged in a specific order. He is physically active and has good endurance. His coordination is intact. His bowel movements are regular. He urinates on his own and asks for a bowel movement. His sleep is undisturbed and he does not wake up at night. He has not had any bedwetting.

[0059] Test results from January 2023: S100 - normal, Neuron-specific enolase 20.6 μg / l, Antinuclear factor normal, CPK, lactate normal, Vitamin D (total) 27.6 ng / ml Blood amino acids: slight increase in taurine, phosphoethanolamine.

[0060] Organic acids in urine - lactate 58 (from here on the measurement is in mmol / mol creatinine), pyruvic acid 43. 2-KG+. Para-hydroxyphenyllactic+. Mandelic is decreased. Suberic. Ethylmalonic, methylmalonic, methylsuccinic are decreased. Kynurenic 0.714. Orotic 2.95. Tricarbyl+.

[0061] The patient underwent a 12-day course of daily procedures: interval hypoxic-hyperoxytherapy, hyperbaric oxygenation, and rectal ozone therapy.

[0062] Intermittent hypoxic-hyperoxytherapy:

[0063] The mode selection was based on the level of immersion in hypoxia and heart rate indicators.

[0064] Interval hypoxia-hyperoxytherapy therapy started with the "D4" mode: 2 minutes of breathing with a mixture of 14% oxygen, 4 minutes of breathing with a mixture of 30% oxygen, 3 minutes of breathing with a mixture of 14% oxygen, 4 minutes of breathing with a mixture of 30% oxygen, 4 minutes of breathing with a mixture of 14% oxygen, 4 minutes of breathing with a mixture of 30% oxygen, 3 minutes of breathing with a mixture of 13% oxygen, 5 minutes of breathing with a mixture of 30% oxygen, 4 minutes of breathing with a mixture of 13% oxygen, 5 minutes of breathing with a mixture of 30% oxygen, 4 minutes of breathing with a mixture of 13% oxygen, 5 minutes of breathing with a mixture of 30% oxygen, 4 minutes of breathing with a mixture of 13% oxygen, 5 minutes of breathing with a mixture of 30% oxygen. The duration of the procedure is 47 minutes.

[0065] Over the course of 12 days, due to the body’s adaptation to hypoxia, the regimes changed towards regimes with an increasingly longer duration of the hypoxia stage with a lower oxygen content in the gas mixture.

[0066] On day 12, the "C1" mode was achieved: 3 minutes of breathing with a mixture containing 14% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen, 4 minutes of breathing with a mixture containing 14% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen, 4 minutes of breathing with a mixture containing 14% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen, 4 minutes of breathing with a mixture containing 14% oxygen, 3 minutes of breathing with a mixture containing 30% oxygen, 5 minutes of breathing with a mixture containing 14% oxygen, 3 minutes of breathing with a mixture containing 30% oxygen, 5 minutes of breathing with a mixture containing 13% oxygen, 3 minutes of breathing with a mixture containing 30% oxygen. The duration of the procedure is 47 minutes.

[0067] Hyperbaric oxygenation was carried out in a pressure chamber with a pressure of 1.3 atmospheres, using a 100% oxygen mixture, a flow of 10 liters per minute using a mask.

[0068] Ozone therapyThe procedure was performed by intravenous drip infusion of ozonated saline with an ozone concentration of 85 μg / ml at a rate of 200 ml per 100 ml of saline solution in a total volume of 300 ml. Ozonation of the saline solution was performed immediately before the ozone therapy procedure, no later than 10-15 minutes before its start.

[0069] During therapy, developmental progress was noted: the appearance of addressed speech in the form of individual words, improved understanding of addressed speech, the emergence of interest in people around, and a decrease in the desire to perform rituals.

[0070] Example 3.

[0071] 4-year-old patient. Diagnosis: Autism spectrum disorder. Pregnancy due to polyhydramnios, diabetes mellitus, hypertension, and edema. Anhydrous interval of 23 hours. Delivery at 38 weeks. Congenital malformation (CMP) 3650 g, height 53 cm. Apgar 8 / 9. Bilateral cephalohematoma at birth. Latched on to the breast after 7 hours. Hyperbilirubinemia, phototherapy was performed. Discharged on the 5th day. Since birth, muscle hypertonicity. Since birth, sleep disturbances; slept only when rocked. Since birth, loose stools up to 10 times a day. Vaccinations according to age up to 2 years. Motor development according to age. Walked at 11 months. At 1 year, he was often sick, but there were no particularly severe infections. Eye contact formed by 3 months, after 5 months it disappeared. From 11 months on, he was constantly rocking and banging his head. By 1 year, he didn't respond to his name. He was babbling. By 1.5 years, he showed stimuli such as spinning, waving his hands in front of his face, sideways glances, and vocalizations. He was markedly hyperactive. He didn't play with other children; if they approached, he ignored them. He would run away from the playground.Stool: loose, undigested. Gradually normalized with gastroenterological treatment. ABA, neuropsychological work, and metabolic support were performed.

[0072] At 4 years of age, the child uses isolated words, sometimes addressed to others: "give," "give back," "go away." He understands spoken language at a basic level. Eye contact is unstable. There is no pointing gesture. He does not fulfill requests. He does not play with other children and ignores his sister. He exhibits self-aggression and aggression when refused. He plays with a train and a tractor. He does not engage in role-playing. He has formal bowel movements 1-3 times a day. Sleep: He falls asleep slowly and does not wake up at night. He awakens at the usual time. He is frequently ill. Examination revealed periorbital cyanosis. Martin-Bell syndrome was excluded. Lysosomal enzymes are normal. The acylcarnitine profile is normal. VEEG with sleep - no epileptic activity.

[0073] Test results (4 years): Antinuclear factor normal, S100 normal, Neuron-specific enolase 24.1 ng / ml, IL-6 normal, TSH, free T3, free T4 normal, CPK 168 U / l, lactate 3.5 mmol / l.

[0074] Blood amino acids: arginine decreased, aspartic acid decreased, glutamic acid decreased, homocysteine ​​decreased.

[0075] Organic acids in urine: lactate 34 (from here on, the measurement is in mmol / mol creatinine), pyruvic acid 32, ACT 8.9, glutaric, adipic, and suberic acid are decreased. Ethylmalonic acid is decreased. Kynurenic acid is decreased. 2-HMA, citric-malic acid 13.3+.

[0076] The patient underwent a 12-day course of daily procedures: interval hypoxic-hyperoxytherapy, hyperbaric oxygenation, and rectal ozone therapy.

[0077] Intermittent hypoxic-hyperoxytherapy:

[0078] The mode selection was based on the level of immersion in hypoxia and heart rate indicators.

[0079] Interval hypoxia-hyperoxytherapy therapy started with the "D2" mode: 2 minutes of breathing with a mixture containing 15% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen, 3 minutes of breathing with a mixture containing 15% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen, 4 minutes of breathing with a mixture containing 15% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen, 4 minutes of breathing with a mixture containing 15% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen, 3 minutes of breathing with a mixture containing 14% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen, 5 minutes of breathing with a mixture containing 14% oxygen, 4 minutes of breathing with a mixture containing 30% oxygen. The duration of the procedure is 45 minutes.

[0080] Over the course of 12 days, due to the body’s adaptation to hypoxia, the regimes changed towards regimes with an increasingly longer duration of the hypoxia stage with a lower oxygen content in the gas mixture.

[0081] On day 12, the "B7" mode was achieved: 5 minutes of breathing with a mixture containing 10% oxygen, 3 minutes of breathing with a mixture containing 30% oxygen, 5 minutes of breathing with a mixture containing 10% oxygen, 3 minutes of breathing with a mixture containing 30% oxygen, 6 minutes of breathing with a mixture containing 10% oxygen, 3 minutes of breathing with a mixture containing 30% oxygen, 6 minutes of breathing with a mixture containing 10% oxygen, 3 minutes of breathing with a mixture containing 30% oxygen, 4 minutes of breathing with a mixture containing 9% oxygen, 3 minutes of breathing with a mixture containing 30% oxygen, 5 minutes of breathing with a mixture containing 9% oxygen, 3 minutes of breathing with a mixture containing 30% oxygen. The duration of the procedure is 49 minutes.

[0082] Hyperbaric oxygenation was carried out in a pressure chamber with a pressure of 1.3 atmospheres, using a 100% oxygen mixture, a flow of 10 liters per minute using a mask.

[0083] Ozone therapy was carried out rectally in a volume of 30 ml of one-time ozone administration.

[0084] During therapy, developmental progress was noted: improvement of vocabulary, improvement of understanding of spoken language, the appearance of elements of role-playing, and the emergence of interest in people around him.

[0085] Bibliography:

[0086] 1. Ajith T.A. Mitochondria-targeted agents: Future perspectives of mitochondrial pharmaceutics in cardiovascular diseases / TA Ajith, TG Jayakumar / / World J Cardiol. - 2014. - Vol.6, No. 10. - P. 1091-1099

[0087] 2. Bastin J. Exposure to resveratrol triggers pharmacological correction of fatty acid utilization in human fatty acid oxidation-deficient fibroblasts / J. Bastin, A. Lopes-Costa, F. Djouadi / / Hum Mol Genet. - 2011. - Vol. 20, no. 10. - P. 2048-2057

[0088] 3. Fischer I, Barak B. Molecular and Therapeutic Aspects of Hyperbaric Oxygen Therapy in Neurological Conditions. Biomolecules. 2020; 10(9): 1247. https: / / doi.org / 10.3390 / biom 10091247

[0089] 4. Prokopov, A. F. (2012). Intermittent Hypoxia and Health: From Evolutionary Aspects to Mitochondria Rejuvenation. In: Xi, L., Serebrovskaya, T. (eds) Intermittent Hypoxia and Human Diseases. Springer, London https: / / doi.org / 10.1007 / 978-1-4471-2906-6_21

[0090] 5. Rybnikova EA, Nalivaeva NN, Zenko MY and Baranova KA (2022) Intermittent Hypoxic Training as an Effective Tool for Increasing the Adaptive Potential, Endurance and Working Capacity of the Brain. Front. Neurosci. 16:941740. doi: 10.3389 / fnins.2022.94174

[0091] 6. Wang H, Shi X, Schenck H, et al. Intermittent Hypoxia Training for Treating Mild Cognitive Impairment: A Pilot Study. American Journal of Alzheimer's Disease & Other Dementias®. 2020;35. doi: 10.1177 / 1533317519896725

[0092] 7. Agadzhanyan N.A., Efimov A.I. Functions of the body under conditions of hypoxia and hypercapnia / / M., Medicine, 1986, 269 p.; Kolchinskaya A.Z. Mechanisms of action of IHT / / Hypoxia Medical J., 1993, No. 1, pp. 5-8.

[0093] 8. Adamyan N.Yu., Karapetyan M.A. Hypoxytherapy - treatment of oxygen deficiency / / - 2018. - T. 58. - No. 3. - pp. 42-49.

[0094] 9. Velskikh E.S., Zvyagina V.I., Uryasyev O.M. Modern concepts of the pathogenesis and approaches to the correction of mitochondrial dysfunction / / Science of the Young - Eruditio Juvenium. - 2016. - No. 1. - P. 104-112.

[0095] 10. Velskikh E.S., Zvyagina V.I., Uryasyev O.M. Modern concepts of the pathogenesis and approaches to the correction of mitochondrial dysfunction / / Science of the Young - Eruditio Juvenium. - 2016. - No. 1. - P. 104-112.

[0096] 11. Belyakov K.M. Dysmetabolic polyneuropathies: clinical and electroneuromyographic diagnostic criteria, pathogenesis, new methods of restorative therapy / / Doctoral dissertation. - 2009.

[0097] 12. Berezovsky V.A., Volobuev M.I. Training to a lack of oxygen as a method of optimizing mental performance. Proc. VII international. symposium. "Ecological and physiological problems of adaptation". Moscow, 1998.

[0098] 13. Bolgaev A.B. et al. TO THE QUESTION OF APPLICATION OF OZONE THERAPY IN MEDICINE / / International Bulletin of Medical Sciences and Clinical Research. - 2023. - Vol. 3. - No. 7. - P. 7-9.

[0099] 14. Davronov L.O., Niyozov Sh.T., Jurabekova A.T. Treatment of encephalomyelitis and myelitis in children with ozone therapy / / Editor-in-chief: Sukiasyan A.A., Cand. of Economic Sciences, Senior Lecturer. - 2015. - P. 190.

[0100] 15. Dubinina E.E. Oxygen metabolism products in the functional activity of cells (life and death, creation and destruction). Physiological and clinical-biochemical aspects / E.E. Dubinina. - St. Petersburg: Publishing house "Medical Press", 2006. - 400 p.

[0101] 16. Karimova L.A. Method of hyperbaric oxygenation in complex treatment / / Educational Bulletin "Consciousness". - 2012. - V. 14. - No. 1. - P. 12-13.

[0102] 17. Kolesnikov A.N., Dubovaya A.V., B2 Naumenko Yu. Prospects for the use of normobaric hypoxytherapy in children (literature review) / / Bulletin of emergency and reconstructive surgery. - 2019. - Vol. 4. - No. 1. - P. 77-80.

[0103] 18. Kulikov A.G. Ozone therapy - an integral part of physiotherapy / / Physiotherapy, balneology and rehabilitation. - 2005. - No. 4. - P. 3-7.

[0104] 19. Levina O.A. et al. Hyperbaric oxygenation in acute diseases and brain injuries. New possibilities. New solutions / / Neurosurgery. - 2014. - No. 4. - P. 9-15.

[0105] 20. Ozone therapy in neurology / A.V. Gustov, S.A. Kotov, K.N. Kontorshchikova, Yu.P. Potekhina. - Nizhny Novgorod: Information center for cooperation "Litera", 1999. - 243 p. - ISBN 5-900915-31-X. - EDN YLRXRZ.

[0106] 21. Cherevaschenko L.A. et al. Ozone therapy and iodine-bromine baths in the correction of cognitive impairment in patients with chronic cerebral ischemia / / Spa medicine. - 2016. - No. 4. - P. 36-41.

[0107] 22. Shilova O.V. Experience of using intermittent normobaric hypoxic therapy in the treatment of neurotic disorders / / Medical news. - 2005. - No. 11. - P. 104-106.

Claims

1. A method of non-pharmacological therapy of autism spectrum disorders in children, including the complex use of interval hypoxic therapy, carried out daily in the form of alternating inhalations of hypoxic (oxygen content from 9 to 16 vol.%) and hyperoxic (oxygen content from 32 to 40 vol.%) mixtures in training modes for a total duration of 45-50 minutes under the control of cardiovascular system parameters, hyperbaric oxygenation, carried out daily in a pressure chamber under a pressure of 1.3 atmospheres of 100% oxygen mixture, a flow of 10 liters per minute, for 45 minutes using a mask, and ozone therapy, carried out by rectal insufflation of an ozone-oxygen mixture with an ozone concentration of 85 μg / ml in a volume of 20-30 ml, for 12 days.

2. The method according to paragraph 1, characterized in that ozone therapy is carried out by intravenous administration of a physiological solution enriched with ozone, with an ozone concentration of 85 μg / ml at a rate of 200 ml per 100 ml of physiological solution.

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