RU2746379C1 - Method of prediction of development of colorectal tumors - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to medicine and concerns a method of predictiion of development of colorectal tumors in patients with gastroenterological pathologies, wherein the method is characterized by the state of parietal mucin of the colonic mucosa being examined by verification of indicator microorganisms, namely bacteria of bifidobacteria, enterobacteria, staphylococci clostridia geni and fungi of candida genus, and upon detection of replacement of bifidobacteria by enterobacteria at a concentration of 10⁶ CFU/g of bioptic material and more in the parietal mucin of the colonic mucosa a high risk of oncotransformation being predicted, upon detection of increased content of bifidobacteria at a concentration of 10⁸ CFU/g of bioptic material and higher with simultaneous presence of staphylococci at a concentration of 10⁴ CFU/g of bioptic material and more a high risk of polyposis being predicted, upon detection of replacement of bifidobacteria by enterobacteria, clostridia and candida at a concentration of 10⁴ CFU/g of bioptic material and higher a high risk of developing an inflammatory reaction of the colonic mucosa being predicted.

EFFECT: invention allows building adequate tactics of managing patients with established signs of qualitative and quantitative state of indicator microorganisms of microbiota of the gastrointestinal tract, urogenital tract and oropharynx in order to personalize treatment and prevent somatic diseases.

1 cl, 10 ex

Description

The invention relates to medicine and relates to a method for predicting the somatic pathology of infectious and non-infectious genesis of a person.

There is a known method for predicting infectious diseases and assessing health status using an immunological blood test by flow cytofluorometry (RF patent No. 2646503), in which the percentage of Th17-lymphocytes is determined and, based on the criterion of compliance with the age norm, the individual level of Th17-lymphocytes in the peripheral blood is detected among individuals with an unknown history and unclear clinical manifestations, a group of healthy individuals and a group of sick individuals.

RF patent No. 2051386 describes a method for predicting infectious diseases in newborns, in which the activity of lysozyme is determined in the cord blood serum of a newborn, and if this indicator is below 40%, the risk of infectious diseases is predicted in the first 7 days of life.

RF patent No. 2413147, dedicated to a method for predicting the incidence of acute infectious diseases in children in the first year of life, is associated with the determination on the first and fifth days after birth of immune status indicators: the absolute number of lymphocytes, carriers of differentiation markers CD3, CD4, CD8, CD 19, CD56, CD95, the indicator of the spontaneous test with nitro blue tetrazolium (NBTsp), the level of the phagocytic indicator (FP), the concentration in the blood serum of immunoglobulins of classes A, M, G (IgA, IgM, IgG) and then the determination of the prediction index (Ipr) using a certain the equations of multiple linear regression, if the IPR does not exceed the value of 3.2, judge the probability of transferring no more than one acute infectious disease in the first year of life; if Ypres is in the range of 3.2-3.7, the occurrence of 2 to 5 acute infectious diseases in the first year of life is likely; if the IPR exceeds the value of 3.7, the probability of occurrence of more than 5 acute infectious diseases in the first year of life is judged.

In RF patent No. 2595863, the essence of the assessment of the state of human health consists in the fact that when predicting the course of an infectious disease, the indicators of the microbiotope of the mucous open cavities are estimated and, on their basis, the degree of disturbance of the microbiotope of the mucous cavities is determined as an indicator of the degree of disturbance of the microbiocenosis of the biotope of the mucous open cavities, depending on the obtained the result is judged on the state of human health and predicts the course of an infectious disease.

In RF patent No. 2556958, the state of mucosal immunity of mucous open cavities is assessed in predicting the course of infectious and inflammatory processes, where, with an established pathogenetic factor, the degree of microbiocenosis disturbance of a particular biotope is recorded using a set of methods for assessing the levels of colonization resistance factors, namely, normal microflora, opportunistic microflora, immunoglobulins - G, M, A, secretory immunoglobulin A and sc component; and depending on the degree of microbiocenosis disturbance, the state of mucosal immunity is assessed and a favorable outcome with eradication of the pathogen or chronicity with persistence of the pathogen is predicted.

Back in the mid-90s, it was found that the characteristics of the human microbiome are under the overwhelming control of paratypical factors, the leading of which is the composition of the nutrient substrate formed by nutrients and metabolites. Genetic and biochemical changes occurring in the human body are manifested by abnormal metabolites. Such changes occur long before the clinically expressed symptoms of the disease. The microbial, as a dynamic system, reacts almost instantly to changes in the nutrient substrate. This means that the qualitative and quantitative characteristics of the microbiome can be used as an integral indicator of the state of the macroorganism, its metabolic processes, reactivity and perception of various factors and environmental conditions without isolating specific genes, metabolites, or biosynthetic products. The modern concept of domestic health care is based on the "4P" principle. This implies that we must anticipate the possibility of the development of a pathological process or diagnose a pathology at the earliest, practically prenosological stage; if possible, prevent or suppress the disease at the very beginning of its development; all treatment and prophylactic measures should be based on the individual characteristics of the patient, preferably focused on unique genetic stigmata. The transition to a new paradigm of health care became possible thanks to the revolutionary scientific and technological achievements of the 21st century - omics technologies, or technologies of a high level of organization. These technologies are based on the assessment of information from large data sets about genes, proteins, metabolites, etc .; suggest the isolation of not only highly specific biomarkers, but also the development of new integral diagnostic criteria. The constituent parts of omics technology are metagenomics, proteomics, and metabolomics. Despite the introduction of these technologies, there is an important problem of their implementation. Reading the result and isolating a specific biomarker from their innumerable variety requires careful verification of the data obtained and "linking" to specific pathological conditions. In this case, the result will still be probabilistic in nature and only determine the risk of a particular event (Microbiocenoses and human health / Ed. By V.A.Aleshkin, S.S.Afanasyev, A.V. Karaulov. M .: Dynasty, 2015).

A known method for predicting the destabilization of the microbiocenosis of the mammalian organism, including the addition of a liquid sample of the internal environment of the organism or the external environment to the culture of the association of microorganisms in a liquid culture medium containing bacteria of the genera Bifidobacterium, Lactobacterium, Enterococcus and Escherichia coli, taken in a ratio of 1011: 1010: 1010: 1010 CFU / l; the absence of destabilization of the microbiocenosis of the mammalian organism under the influence of the internal environment of the organism or the external environment, respectively, in the absence of a decrease of at least 100 times the concentration of one of the components of the association of microorganisms (RF patent No. 2377311).

However, the known method does not apply to predicting prenosological manifestations of somatic diseases of infectious and non-infectious genesis. It also lacks a personalized forecasting approach.

As a prototype, the textbook Parietal microbiocenosis of the human gastrointestinal tract was chosen (EA Bogdanova, Yu.V. Nesvizhsky, VV Zverev Parietal microbiocenosis of the human gastrointestinal tract (textbook). - M .: Publishing house "Russian doctor ". - 2009), containing information of an informative nature only about the types of microorganisms detected in biotopes of parietal mucin of the large intestine in diverticulosis and polyposis, tumor, inflammation. Also, there is no information on an individual approach to the diagnosis of the above pathologies of the large intestine by the levels of indicator strains in the foci of the development of pathological changes at the prenosological level.

The technical problem to be solved when creating the invention is the development of a method for predicting the somatic pathology of infectious and non-infectious genesis, which will detect deviations in the state of human health at an early or prenosological stage and thereby prevent the onset or deterioration of the course of the disease, decrease in life and work capacity.

The technical result of the present invention is the fact that:

- the claimed method for predicting somatic pathology of infectious and non-infectious genesis allows forecasting at the prenosological stage;

- to evaluate the effectiveness of the therapy;

- monitor the adaptive capabilities of the macroorganism, predict the preservation of life and performance in ecologically unfavorable and critical conditions;

- to identify and register undocumented effects of chemical compounds for medical, household and economic purposes;

- to form an adequate tactics for managing patients with established indicators of the qualitative and quantitative state of indicator microorganisms of the microbiota of the gastrointestinal tract, urogenital tract and oropharynx in order to personalize the treatment and prevention of somatic diseases.

The essence of the invention lies in the fact that patients study the state of the microbiota of the mucous open cavities by verifying the qualitative and quantitative characteristics of indicator strains of microorganisms responsible for molecular genetic abnormalities in the state of the macroorganism. Deviations from the critical values of the levels of qualitative and quantitative indicators of the present indicator strains of microorganisms of the patient indicate the likelihood of the risk of developing a particular pathological condition.

The human body is a complex phylogenetically and ontogenetically formed symbiosis "macroorganism-microbe" with certain relationships. It was previously found that the characteristics of the human microbiome are under the overwhelming control of paratypical factors, the leading of which is the composition of the nutrient substrate formed by nutrients and metabolites. This means that a microbial is a dynamic system, and its characteristics can be used as a universal integral indicator of the state of a macroorganism, its metabolic processes, reactivity and perception of various factors and environmental conditions without isolating specific genes, metabolites, or biosynthetic products. In the human body, as a result of non-lethal mutation in the "readable" region or activation of repressed genes, as well as the onset of biosynthesis of an abnormal protein isoform or certain disorders in the chain of biochemical transformations under the influence of factors of various nature, incl. drugs or toxicants, abnormal metabolites appear that are secreted into the lumen or secretion of mucous hollow organs. Metabolic changes occur long before the clinically expressed manifestations of the disease. The microbiota inhabiting the lumen and parietal mucin of the mucous hollow organs reacts to the appearance of metabolites atypical for a particular person by changing their qualitative and quantitative characteristics: the appearance or disappearance of their typical representatives or a pronounced change in their content. Qualitative and quantitative changes in the human microbiota are indicators of the unfavorable state of the human body at the prenosological stage and indicate the likelihood of developing certain diseases and conditions.

In this regard, it is necessary to develop integral indicators of molecular genetic shifts in the human body, which would make it possible to detect deviations in the state of human health at an early or prenosological stage, as well as the adaptive capabilities of his body, and thereby prevent the onset or worsening of the course of the disease, decrease in life. - and efficiency.

The adopted concept made it possible to take a fresh look at the results of microbiological studies of patients and to identify candidate biomarkers of certain conditions. The following examples illustrate our thesis well:

- in the study of parietal mucin of the mucous membrane of the large intestine of patients with gastroenterological pathologies in the zone of oncotransformation, bifidobacteria are replaced by enterobacteria (at a concentration of 10 ³ CFU / g and above biopsy); in the polyp area, an increased content of bifidobacteria 10 ⁸ CFU / g and above biopsy specimen and staphylococci 10 ³ CFU / g and above biopsy specimen is recorded; in the area of inflammation, bifidobacteria (not sown) are replaced by enterobacteria, clostridia and candida in a concentration in all cases of 10 ³ CFU / g and above biopsy;

- according to the results of a study of the microbiota of feces in subacute chronic endocarditis, the presence of lactose-negative Escherichia coli and Staphyloccus aureus is recorded - both at a concentration of 10 ³ CFU / g and higher;

- in rheumatism - the presence of E. coli with reduced enzymatic activity at a concentration of 10 ³ CFU / g and higher and the absence of non-pathogenic staphylococci (not sown) is recorded;

- in chronic renal failure - the presence of lactose-negative E. coli and E. coli with hemolytic activity is recorded - both at a concentration of 10 ³ CFU / g and higher;

- in the study of the fecal biotope of patients with diabetes mellitus, sowing of the yeast-like fungus Saccharomyces cerevisae at a concentration of 10 ³ CFU / g and higher in persons under 50 years of age (especially without candidiasis in history) indicates a violation of carbohydrate metabolism and, most often, type 2 diabetes mellitus ; in patients with more than 5 years of illness, a high risk of developing late complications of diabetes mellitus;

- in a comprehensive study of the microbiota of pregnant women in patients with a compensated threat of termination of pregnancy, clostridia are verified in feces at a concentration of 10 ⁷ CFU / g and above, and in the oropharynx - peptostreptococci at a concentration of 10 ⁴ CFU / g and above; if it is impossible to compensate for the threat of miscarriage in the oropharynx - S. aureus at a concentration of 10 ⁴ CFU / g and above, and in the vagina - peptostreptococci and coagulase-negative staphylococci, both at a concentration of 10 ⁴ CFU / g and above.

In all cases, the discriminatory efficiency of indicator biomarkers reached 75%.

The claimed method for predicting somatic pathology of infectious and non-infectious genesis is new and has not been described in the literature.

As evidence of the possibility of realizing the declared purpose and achieving the specified technical result, we present the following data.

Clinical example 1

Patient A., pregnancy 8-9 weeks. During the visit to the gynecologist of the antenatal clinic, a sterile brush was used to take swabs-scrapings from the mucous membrane of the vagina, oropharynx, as well as from the lumen of the rectum, and a microbiological study was carried out. The smear-oropharyngeal mucosa scrapings from peptostreptokokki detected at concentrations of 10 ⁵ cfu / g in feces - clostridia in a concentration of 10 ⁸ cfu / g.

The possibility of the development of spontaneous abortion, which can be compensated by medication, was suggested.

At 11-12 weeks, the patient was at risk of spontaneous abortion. Treatment was prescribed.

Outcome: the threat of spontaneous abortion is compensated, treatment is effective, urgent uncomplicated childbirth.

Clinical example 2

Patient B., pregnancy 10-12 weeks. During the visit to the gynecologist of the antenatal clinic, a sterile brush was used to take swabs-scrapings from the mucous membrane of the vagina, oropharynx, as well as from the lumen of the rectum, and a microbiological study was carried out. Staphylococcus aureus was found in a scraping smear from the mucous membrane of the oropharynx at a concentration of 10 ⁵ CFU / g, peptostreptococci and coagulase-negative staphylococci were isolated from the walls of the vagina, both at a concentration of 10 ⁴ CFU / g.

The possibility of developing incurable spontaneous abortion was suggested.

At 13-14 weeks, the patient was at risk of spontaneous abortion. Treatment was prescribed.

Outcome: the threat of spontaneous abortion could not be compensated for, spontaneous abortion at 14-15 weeks.

Clinical case 3

Patient V., 32 years old. With a periodic medical examination, he does not show any complaints about his well-being. He denies a history of candidiasis episodes.

The results of a biochemical blood test are within normal limits.

During microbiological examination of fecal contents, the yeast-like fungus Saccharomyces cerevisae was sown at a concentration of 10 ⁴ CFU / g.

It was suggested that the patient might develop a disorder of carbohydrate metabolism.

At the next periodic medical examination after 1 year, the patient complains of increased sweating, weight gain, increased thirst, anxious sleep, and increased fatigue.

Fasting glucose (sugar) level -7.1.

In the microbiological study of fecal contents, the yeast-like fungus Saccharomyces cerevisae continues to be sown at a concentration of 10 ^3-5 CFU / g.

Diagnosis: type 2 diabetes mellitus. Treatment was prescribed.

Clinical case 4

Patient G., 52 years old. Diabetes mellitus type 2 for 9 years.

During microbiological examination of fecal contents, the yeast-like fungus Saccharomyces cerevisae was sown at a concentration of 10 ⁴ CFU / g.

Conclusion: high risk of developing late complications of diabetes mellitus.

Outcome: Diabetic nephropathy developed within 3 years. Treatment was prescribed.

Clinical case 5

Patient D., 67 years old. During a medical examination in a hospital, complaints of unstable stools. To exclude pathology of the large intestine, it was proposed to undergo a colonoscopic examination.

During colonoscopy, a mucosal biopsy is taken. The biopsy specimen was cultured. In the course of a microbiological study, an increased content of bifidobacteria was found at a concentration of 10 ⁹ CFU / g of biopsy and staphylococci at a concentration of 10 ⁶ CFU / g of biopsy.

The possibility of developing colon polyposis was suggested.

On repeated medical examination after 1 year complaints of unstable stools, recurrent bloody discharge from the rectum.

During colonoscopy, a polyp was found, and a biopsy was taken. A histological examination was carried out.

Diagnosis: adenomatous polyp. Treatment was prescribed.

Clinical case 6

Patient D., 42 years old. With a periodic medical examination, he does not show any complaints about his well-being. The mother has a history of a colon polyp. Colonoscopy recommended.

During colonoscopy, a mucosal biopsy is taken. The biopsy specimen was cultured. During microbiological investigations revealed the complete absence of bifidobacteria contained in a concentration of enterobacteria June ¹⁰ CFU / g biopsy.

Suspicion of the possibility of developing a neoplastic process.

On repeated medical examination 1 year later, complaints of unstable stools, periodic bloody discharge from the rectum appeared. Colonoscopy was scheduled.

During colonoscopy, a suspicious area of a neoplastic process up to 2 cm in size was found, a biopsy was taken. A histological examination was carried out.

Diagnosis: Histological confirmation - TisN0M0 colorectal cancer. Treatment was prescribed.

Clinical case 7

Patient J., 35 years old. With a periodic medical examination, he does not show any complaints about his well-being. The mother has a history of colorectal cancer. Colonoscopy recommended.

During colonoscopy, a mucosal biopsy is taken. The biopsy specimen was cultured. During the microbiological study, a complete absence of bifidobacteria was found, enterobacteria were contained in a concentration of 10 ⁷ CFU / g of biopsy, Clostridia and Candida - 10 ⁴ CFU / g of biopsy.

Suspicion of the possibility of developing an inflammatory reaction.

On repeated medical examination 1 year later, complaints of unstable stools, periodic bloody discharge from the rectum appeared. Colonoscopy was scheduled.

During colonoscopy, large areas of inflamed mucosa were found, and a biopsy was taken. A histological examination was carried out.

Diagnosis: Non-ulcerative colitis. Treatment was prescribed.

Clinical case 8

Patient I., 27 years old. With a periodic medical examination, complaints of easy fatigue, weakness, periodic nausea and a slight rise in body temperature to subfebrile values.

The results of a biochemical blood test were within normal limits; physical examination did not reveal any signs of pathology.

To exclude intestinal dysbiosis, a microbiological study was prescribed.

Microbiological examination of fecal contents showed lactose-negative Escherichia coli at a concentration of 10 ⁴ CFU / g and Staphylococcus aureus at a concentration of 10 ⁵ CFU / g.

The possibility of developing infective endocarditis in the patient was suggested.

Within 6 months. the patient developed discomfort in the region of the heart, stable subfebrile condition with periodic sharp rises in temperature up to 38.6-39.0 ° C, sweating, weakness, muscle pain. Examination of the skin - petechial rash.

Suspected subacute infective endocarditis. Hospitalized.

Inpatient examination:

Electrocardiography is an instability of the heart rhythm.

Radiography - an increase in the boundaries of the heart.

General and biochemical blood tests: blood color index - 0.75, erythrocyte level - 3.5 × 10 ¹² cells / l of blood, leukocyte level - 10 ¹⁰ cells / l, ESR - 62 mm / h, an increase in the amount of gamma globulins and alpha ₂ -globulins of blood plasma.

Bacteriological examination of blood at the peak of fever - Staphylococcus aureus was sown.

Diagnosis: subacute infective endocarditis. Treatment was prescribed.

Clinical case 9

Patient K., 39 years old. With a periodic medical examination, complaints of easy fatigue, weakness, periodic nausea and a slight rise in body temperature to subfebrile values.

The results of a biochemical blood test were within normal limits; physical examination did not reveal any signs of pathology.

To exclude intestinal dysbiosis, a microbiological study was prescribed.

In a microbiological study of fecal contents, E. coli with reduced enzymatic activity at a concentration of 10 ⁴ CFU / g was inoculated, non-pathogenic staphylococci were completely absent.

The possibility of developing rheumatism in the patient was suggested.

Within 8 months. the patient developed discomfort in the region of the heart, persistent subfebrile condition with periodic sharp rises in temperature up to 38.6-39.0 ° C, sweating, weakness, muscle pain, joint aches. Examination of the skin - pale, petechial rash.

Suspicion of acute rheumatism. Hospitalized.

Inpatient examination:

Electrocardiography is an instability of the heart rhythm.

Radiography - an increase in the boundaries of the heart.

General and biochemical blood tests: the level of neutrophils - 2 × 10 ¹⁰ cells / l, ESR - 62 mm / h, an increase in the amount of gamma globulins, alpha ₂ -globulins and blood plasma fibrinogen, antistreptolysin-O titer - 2500 units, CRP - 4.2 mg / l, rheumatoid factor - 28 IU / l.

Diagnosis: acute rheumatism. Treatment was prescribed.

Clinical case 10

Patient L., 55 years old. Urolithiasis for 7 years.

In a microbiological study of fecal contents, E. coli with hemolytic activity and lactose-negative E. coli were seeded - both at a concentration of 10 ⁴ CFU / g.

The possibility of developing chronic renal failure was suggested.

Outcome: clinical and laboratory signs of chronic renal failure developed within 2 years.

The proposed method for predicting the development of deviations in the state of human health at an early or prenosological stage by determining integral indicators of molecular genetic shifts in the human body, as well as an algorithm for managing patients with prenosological pathological manifestations of diseases, which determines the choice of adequate therapeutic and prophylactic measures depending on the verification and state indicator microflora of the microbiota of the mucous open cavities of the body and the severity of prodromal symptoms are modern trends in domestic medicine in preventing the onset or worsening of the course of the disease, reducing life and working capacity.

Claims (1)

A method for predicting the development of colon neoplasms in patients with gastroenterological pathologies, characterized by the fact that the state of parietal mucin of the colon mucosa is investigated by verification of indicator microorganisms, namely bacteria of the genus Bifidobacteria, enterobacteria, staphylococci, clostridia and fungi of the genus Candida, and when detecting enterobacteriaceae at a concentration of 10 ⁶ CFU / g of biopsy and more in the parietal mucin of the colon mucosa, a high risk of oncotransformation is predicted; if an increased content of bifidobacteria is detected at a concentration of 10 ⁸ CFU / g of biopsy and higher with the simultaneous presence of staphylococci at a concentration of 10 ⁴ CFU / g of biopsy or more, a high risk of polyposis is predicted; when detecting replacement of bifidobacteria by enterobacteria, clostridia and candida at a concentration of 10 ⁴ CFU / g of biopsy and higher, a high risk of developing an inflammatory reaction of the colon mucosa is predicted.