Method for increasing regeneration activity of rat's intestinal epithelium following radiation exposure

FIELD: medicine.

SUBSTANCE: allogenic multipotent mesenchymal stromal cells in the number of 6 million cells/kg are introduced intravenously in a dose of 3.0 Gy into rats suffered from radiation exposure. Additionally, haemopoietic stem cells recovered from umbilical blood in the number of 300 thousand cells/kg are introduced. The cells are transplanted in such a manner 60 min after radiation exposure.

EFFECT: higher mitosis count that leads to a substantial cryptal epithelium increase.

3 tbl

 

The invention relates to experimental medicine, namely to develop ways to treat radiation sickness, and can be used to restore the epithelium of the jejunum after exposure to radiation.

The nature of radiation damage to the body of the animal and the person intensively studied for several decades. Gastro-intestinal tract (GIT) refers to critical systems cell renewal during irradiation. Under the action of ionizing radiation (AI) (radial load) in the mucosa of the digestive tract sharp violations of dynamic equilibrium between individual pools, leading to severe functional disorders in the system. Depending on the severity of the disorders that may be essential for the functioning of the entire organism. Supplementation of cell loss is a top priority of the ongoing functional therapy, contributing to the weakening of the primary disorders that threaten life. Primary violation in the small intestine is a cellular devastation of the villi and crypts.

Recovery regeneration cryptologi epithelium after acute radiation damage can be reduced to the proliferation of cells retained viability, due to which compensated the decline in the population of cells, and thus restored their functionality is Naya completeness (I. Vasilenko. Radiation ecology [Text] / Vasilenko I. M.: Medicine, 2004. - 88 C.).

The next direction in the pathogenetic therapy of radiation lesions of the intestinal epithelium is the use of metabolic resources (GL the vishkovsky. Register of medicines of Russia (Text) / GL the vishkovsky. - M.: radar, 2005. - S).

Drug - bottled hydroxytrol can be used in radiation lesions of the mucous membrane of the intestine, as has expressed an antioxidant, antihypoxic, anti-inflammatory, regenerating effect.

The disadvantages of the application of these products (metabolic funds and, in particular, bottled hydroxytoluene) are side effects such as allergic reactions, urethritis, prostatitis.

Restoring the regeneration of the epithelium of the jejunum is provided by the use of drugs that increase the regenerative potential of the epithelial cells of the crypts. A typical representative of drugs that have a stimulating effect on the regeneration processes is Derinat. Derinat is a sodium salt of DNA obtained from sturgeon milk. This drug is used not later than 24 h after irradiation. It is injected once intramuscularly or subcutaneously in a volume of 15 ml (75 g of active substance) (Kutsenko S.A. Military toxicology, radiobiology and medical is nska protection [Text]: textbook. a manual for students of honey. universities / S. Kutsenko, NV, Butoma, A.N. Grebenyuk. - SPb.: Folio, 2004. - 527 S.).

However, the use of this group of drugs is associated with side effects and contraindications. Thus, after injection of an aqueous solution of Derinat in 1.5-3 h there is a severe hypoglycemia (GL the vishkovsky. Register of medicines of Russia (Text) / GL the vishkovsky. - M.: radar, 2005. - S-304).

The closest solution to the claimed method is the recovery of the epithelium of the jejunum laboratory animals after exposure to ionizing radiation, which is carried out in 25-30 min after irradiation by intravenous transplantation of allogeneic multipotent mesenchymally stromal cells (MMSC) in the amount of 6·106cells/kg (RU, patent No. 2415476, IPC G09B 23/28, AK 35/50, OR 43/00, publ. 27.03.2011,) prototype.

This invention allows to expand the Arsenal of tools capable of providing regenerative potential of tissues, namely antiapoptotic action, the increase in the rate of cell renewal cryptologi epithelium of the jejunum, oppressed by ionizing radiation, in the absence of complications, however, there are theoretical reasons to suppose that the use of combined transplantation MSC and hematopoietic stem cells (HSCs) could provide more virginmegastore to activate the regeneration of myeloid tissue in comparison with the introduction of only MMSC.

Object of the invention is the expansion of the means capable of providing regenerative potential of tissues.

The technical result that will be achieved from the use of the invention is to increase the activation of the regeneration of the intestinal epithelium.

The technical result is achieved in that in the method of restoring the regeneration of the epithelium of the intestine of laboratory animals after radiation exposure by intravenous transplantation of allogeneic multipotent mesenchymally stromal cells in the amount of 6 million cells/kg impose additional hematopoietic stem cells (HSC) in the amount of 300 thousand cells/kg, the cell transplantation is carried out after 60 min after radiation exposure.

The invention consists in the application for the first time to restore the regenerative capacity of the epithelium of the intestine combined stem cell transplantation: MMSC and GSK, selected, respectively, from the placenta and umbilical cord blood.

It is known that MSC, producing a chemoattractant for HSC (SDF-1), regulate HSC homing, enhancing the migration of the latter in the place of the most damaging tissue. MMSC produce the whole spectrum of anti-inflammatory cytokines (IL-4, 10, TGF-β) and are able to exert a cytoprotective effect inducyruya the production of heat shock proteins (HSP70) is located near the s cells, as well as producing the cytokine HIF-1α, can inhibit apoptosis. The indicated cytokine profile may lead to changes in cell population, to reduce the severity of the inflammatory response and reduce the damage zone. MSC able to migrate to the injury site, to gain a foothold, differentiation, and function as substituted by one. Intravenous transplantation of allogeneic MSC in the amount of 6 million cells/kg after 60 minutes after irradiation does not occur reactions, transplant rejection, and does not occur graft versus host due to the immunosuppressive properties MMSC. These properties MMSC give the opportunity to use them to restore the regeneration of intestinal epithelial cells. Eksperimentalno proven ability to merge GSK in the amount of 300 thousand cells/kg with the stem cells of the intestine, which then differentiate into Mature enterocytes. This mechanism contributes to the activation of the regeneration of the intestinal epithelium.

From the analysis of scientific-technical and patent information, the use of combined transplantation MSC and GSK in the inventive quantitative limits, isolated from the placenta, allowing after exposure to ionizing radiation to increase the mitotic activity of the cells, which leads to a significant increase in the content cryptologi epithelium, and SL is therefore to restore the regeneration of the intestinal epithelium, we have not identified that allows to make a conclusion on the conformity of the proposed technical solution the criteria of "novelty" and "inventive step".

The invention is carried out as follows.

General characteristics of laboratory animals used in research

The experiments were performed on 42 white laboratory rats male age 6-8 months, weighing 200-220 g Experiments to obtain MSC and GSK performed on 9 laboratory animals the female-female rats aged 3-4 months, weighing 150-170 g, gestation 18 days. The animals were kept in standard laboratory animal facility under the Rules of work with the use of experimental animals"approved by Order of the USSR Ministry of health No. 755 from 12.08.1977, and the Order of the USSR Ministry of health No. 1179 from 10.10.1983 "On approval of regulations of the cost of feed for laboratory animals in health care facilities".

The number of experimental animals and their distribution on the series of experiments presented in table 1.

Have investigated the effects of ionizing radiation dose of 3.0 G on laboratory animals Mature age, were allocated to experimental and control groups (without the introduction of MSC and GSK). Animals of the experimental group was injected intravenously suspension MMSC and GSK respectively at a dose of 6 million cells/kg and 300 is is. cells/kg in 0.5 ml of 0.9% NaCl. Animals of the control group was injected intravenously with 0.5 ml of 0.9% NaCl. There have also been allocated intact laboratory animals (not exposed), which was injected intravenously 0.5 ml of 0.9% NaCl. Intravenous administration was carried out after 60 minutes after irradiation once at the above doses. The slaughter of animals was carried out at 1 and 7 days after irradiation (table 1).

Table 1
The distribution of animals on the series of experiments after exposure to ionizing radiation dose of 3.0 G
Laboratory animals
The autopsy of the bodiesThe placeholderThe autopsy of the bodies
The inventive method
24 hoursday 724 hoursday 7
AI, a dose of 3.0 GExperienced groupIntravenous MSC 6 million cells/kg and HSC 300 thousand cells/kg in 0.5 ml 0.9% NaCl7 PCs
7 PCs7 PCsIntravenous MSC 6 million cells/kg in 0.5 ml 0.9% NaCl7 PCs
The control group
Intravenously 0.5 ml of 0.9% NaCl 7 PCs7 PCsIntravenously 0.5 ml of 0.9% NaCl7 PCs7 PCs
Without the impact of AIThe intact
Intravenous
0.5 ml of 0.9% NaCl
7 PCs7 PCsIntravenously 0.5 ml of 0.9% NaCl7 PCs7 PCs

Assessment methods regenerative processes in the mucosa of the jejunum

To study the overview histologic and morphometric analysis in all series of experiments for the study was carried autopsy jejunum.

The operation was performed under ether anesthesia. On average the line of the anterior abdominal wall was made the cut length of 3 cm, removed the loops of the small intestine. Resection was exposed to the upper third of the jejunum.

Given the presence of circadian rhythms, the animals were killed by decapitate at the same time of day (9-10 am). The operation was conducted in the first half of the day.

The material was fixed in 10% neutral formalin. The fill was performed in paraffin and prepared slices with strict orientation of the villi and crypts in the thickness of 5 μm, which were stained with hematoxylin and eosin and van gieson.

- Assessed the mitotic activity of the intestinal epithelium by determining the mitotic index (MI) on histological sections of the longitudinal orientation. Counted the epithelial cells with the four stages of mitosis meet at 3000-3500 cells, the result expressed as a percentage.

MAnd=Kaboutlandhewith atinaboutmitotically dividing epithelial cells3000-3500it is estimated epithelial cells of the crypts*100%

- Rated programmed death of epithelial cells by determination of apoptotic index (AI) on histological sections of the longitudinal orientation. Calculated epithelial cells with morphologically identified to the chinoy apoptosis meet at 3000 cells, the result expressed as a percentage.

AndAnd=Kaboutlandhewith atinaboutepithelial cells in apoptosis3000it is estimated epithelial cells of the crypts*100%

- Average cellularity in one crypt was determined as the ratio of the total number counted kaptelinin cells to the number of the analyzed crypts, expressed in percent (Trufakin VA Methods for evaluating the proliferation and differentiation of the intestinal epithelium. Methodical recommendations, Novosibirsk, 1990). Histological preparations of the jejunum were analyzed using microscope Micros MS - 50 (Austria) at magnification 100*15.

Morphology and quantification of apoptosis was performed with fluorescence microscopy using two fluorochromes: acridine orange (made in Finland) and Annexin V - FITC fluorescence microscopy (U.S. production).

Culture MMSC

Produced highlighting the placenta (gestation 18 days), a tissue sample weight of 1 g three times washed with physiological saline, phosphate buffered (PBS) at pH 7.2, without ions of CA2+and Mg2+,supplemented with antibiotics (penici the Lin 50 u/ml, streptomycin 50 μg/ml), then followed by mechanical grinding tissue and enzymatic treatment (0.25% trypsin - EDTA for 15 min at 37°C). The resulting cell suspension was filtered twice through a 100 μm nylon membrane for removal of large unground pieces of fabric. The suspension was diluted with medium α-MEM containing antibiotics, the cells are then besieged by centrifugation for 15 minutes at 1000 g. The obtained cell sediment suspended in the medium α-MEM (ICN, USA) with 10% serum of cow embryos (Hyclone, New Zealand), a single solution of essential amino acids (Sigma, USA) and a single antibiotic solution (Chemicon). Suspension cells were sown at a concentration of 150000-160000 CL/cm2on Petri dishes 60 mm (Nunc, Denmark).

1.1. The cultivation conditions MMSC

The cultivation was performed at 37°C in humidified atmosphere with 5% CO2.

1.2. Counting cells

Counting cells produced in hemocytometer. Calculate the number of cells in 1 ml of the suspension was made according to the formula X=a*10000, where X is the number of cells in 1 ml of suspension; and - the amount of cells in small squares.

1.3. Subcultivation MMSC

Subcultivation cells was carried out based on the achievement of 80% of the monolayer. Cells were washed with a mixture of 0.25% trypsin - EDTA at a ratio of 1:1, in which the cells are incubated for about 5 min at 37°C. Trypsin iactiveaware addition ro the postal environment with CEQ (10%) and the suspension was centrifuged at 200 g for 5 minutes After removing the supernatant, the cells are suspended, made calculations and were sown in the desired concentration (10000 cells/cm2).

1.4. The characteristics of the culture was performed using a set of primary Mesenchymal Stem Cell Characterization Kit) and secondary antibody (secondary antibody Cy3 conjugated).

Selection GSK

To highlight GSK has used a set of Mouse Hematopoietic Progenitor (Stem) Cell Enrichment Set-DM (StemCell Technologies, Canada).

Methods of statistical processing of the obtained results

Each row of the metric value was calculated arithmetic mean, standard error of the mean. The reliability of differences between the prototype and the application was evaluated using a t-student criterion. Differences were considered significant at p<0,05.

Indicators regenerative activity of the jejunal epithelium of laboratory animals at 1 day after exposure to ionizing radiation dose of 3.0 G, M±m, n=7 are shown in Table 2.

Table 2
Application (introduction MSC and GSK)Intact animals
IndexThe control groupPrototype (MMSC)
Mitotic index, %7,33±1,12°*
5,73±0,86°4,82±1,16°8,88±0,74
Apoptotic index %13,85±1,32°8,01±0,23°*
12,77±1,71°and 4.68±0,41
Average cellularity 1 of the crypt (CCMS)
55,82±3,74°49,20±4,80°64,24±3,15°*73,22±4.26 deaths
Note: *the difference values in the experimental gr is the group in the application of indicators in the experimental group in the prototype, significantly with p<0,05,
° the difference values from the parameters of the intact animals, significantly with a p<0,05.

It should be noted that on the 1st day after introduction MMSC on the impact of AI in laboratory animals indicators regenerative activity of the epithelium of the jejunum did not significantly differ from values in the control group. At the same time after combined transplantation MSC and GSK when counting the number of mitoses in the epithelial cells of the crypts revealed that this indicator in the application 52.1% higher than in the prototype. When counting the number of cells in apoptosis was established that the studied parameter in the application by 42.2% lower than in the prototype. When calculating the average cellularity 1 of the crypt established that the studied parameter was 30.1% higher than in the prototype.

Thus, the obtained data indicate that co-transplantation MSC and HSC on the 1st day after exposure to ionizing radiation through the increase in the number of mitoses and through the inhibition of apoptosis leads to the increase of the content of cryptologi epithelium significantly greater extent than the introduction of only MMSC.

Indicators regenerative activity of the jejunal epithelium of laboratory animals at 7 days after exposure to ionizing radiation dose of 3.0 G, M±m, n=7 are shown in Table 3.

Table 3
Application (introduction MSC and GSK)Intact animals
IndexThe control groupPrototype (MMSC)
Experienced group
Experienced group
Mitotic index, %12,08±0,92*
of 8.92±0,9216,23±3,21***8,88±0,74
Apoptotic index %
6,60±0,63°4,75±0,72*a 4.83±0,77*and 4.68±0,41
Average cellularity 1 of the crypt (CCMS)
72,82±was 4.0288,15±4,42*106,73±5,68***73,22±4.26 deaths
Note: * difference from control group animals exposed to AI dose of 3.0 G, significantly with a p<0,05; ** difference values in the experimental group in the application of indicators in the experimental group in the prototype, significantly with a p<0,05;° the difference values from the parameters of the intact animals, significantly with a p<0,05.

As can be seen from table 3, in laboratory animals, which was produced by intravenous transplantation MSC on the 7th day after exposure to AI when counting the number of mitoses in the epithelial cells of the crypts found that this rate was 34.4% more than in the prototype, and did not differ doscover is from intact animals. Counting of cells in apoptosis was established that the content of these cells was not significantly different from the same indicator as in the prototype, and the value of intact animals. When calculating the average cellularity 1 of the crypt is found that the studied parameter in the request was 21.1% higher compared with the value in the prototype and did not significantly differ from the values in the intact animal.

Thus, by the 7th day after exposure to ionizing radiation is a natural recovery of the number of mitoses in the epithelial cells of the crypts and content cryptologi epithelium (figure CCM). At the same time, evidence suggests that co-transplantation MSC and HSC on the 7th day after exposure to ionizing radiation through the increase in the number of mitoses leads to an increase in the content cryptologi epithelium significantly greater extent than the introduction of only MMSC.

The way to increase the regenerative activity of the intestinal epithelium of rats after radiation exposure at a dose of 3.0 Grams, including intravenous allogeneic multipotent mesenchymally stromal cells isolated from the placenta, in the amount of 6 million cells/kg, characterized in that it further injected hematopoietic stem cells isolated from umbilical cord blood, in the amount of 300 thousand cells/kg, and akuu the cell transplantation is carried out after 60 min after radiation exposure.



 

Same patents:

FIELD: medicine.

SUBSTANCE: method involves simulating endothelial dysfunction in Wistar male rats by daily intraperitoneal introduction of L-nitro-arginine-methyl ester 25 mg/kg/day for 7 days. Endothelial dysfunction is corrected by intragastric introduction of an api-phytocomposition in the form of an aqueous suspension of honey, royal jelly, bee bread, Resveratrol in the ratio of 10:1:1:2. The composition is introduced in a dose of 200 mg/kg of an animal's body weight once a day for 7 days.

EFFECT: method provides synergetic endothelioprotective effects with no side effects observed when treating with synthetic preparations.

1 ex

FIELD: medicine.

SUBSTANCE: method involves simulating hepatitis by 7-day alcoholisation in female rats. A hepatoprotective agent is presented by an api-phytocomposition in the form of an aqueous suspension of honey, lecithin and licorice root extract in the ratio of 10:2:1. The composition is introduced twice a day intragastrically before feeding in a dose of 200 mg/kg of body weight 5 days before ethanol introduction and for 7 days one hour before ethanol introduction.

EFFECT: effective treatment of hepatitis with no side effects.

1 ex, 1 tbl, 1 dwg

FIELD: medicine.

SUBSTANCE: method involves a supramedian laparotomy, providing an approach to a lower one-third of the oesophagus and introducing benzalkonium chloride into the gastro-esophageal junction. The above agent is injected in the concentration of 0.02% into 4 points at 3, 6, 9 and 12 o'clock in a projection of the cardiac sphincter in an amount of 0.2-0.3 ml in each injection point.

EFFECT: higher clinical effectiveness.

8 dwg

FIELD: medicine.

SUBSTANCE: selenomethionine in the concentration of 2 mcg/ml in a single dose of 10 mcg/kg of body weight is administered intraperitoneally into an experimental animal. One day later, surgical anaesthesia involves a standard fracture of the upper one-third diaphysis of the femur with an intramedullary osteosythesis with a steel nail.

EFFECT: method enables providing the bone resorption in the process of osteogenesis with full reproducibility.

6 dwg

FIELD: medicine.

SUBSTANCE: balanced ration to be gradually changed by cholesterol and pork fat is fed in male rats aged more than one year. Vitamin D2 in an amount of 35,000 IU/kg to 60,000 IU/kg of body weight is orally introduced. The above manipulations are carried out with an underlying exposure to stress factors: noise, varying photoperiod and periodic hypoxia.

EFFECT: method provides inducing pathoanatomical, haematological, biochemical and histological changes specific for atherosclerosis and close to those in a human.

4 ex, 2 tbl

FIELD: medicine.

SUBSTANCE: method is implemented by applying an innovative control method which involves both instrumental and visual control methods by specific parameters, as well as training for basic cardiopulmonary resuscitation in children and adolescents with using stimulation technologies. The given method possesses high availability of the doctor's theoretical knowledge and skills. The method involves controlling the cardiopulmonary resuscitation skills by doctors before the cardiopulmonary resuscitation training, theoretical and practical training, as well as diagnosing the achieved level of cardiopulmonary resuscitation. A trainer conducts control by means of a developed point rating control system and using a training manikin. Upon termination of control, the individual doctor rating is calculated.

EFFECT: higher survival rate of the children with cardiac arrest.

4 tbl

FIELD: medicine.

SUBSTANCE: external fixation apparatus (EFA) is applied on a rabbit's shin, then an osteoperforation and an osteoclasis are performed between the 2nd and 4th levels. Shin bone fragments are separated at 1 cm and fixed with EFA pins in this position for 14 days; that is followed by a one-stage compression between supports of the 4th and 5th levels until resist completely. Ends of the separated fragments are combined and fixed; the EFA is stabilised. A device for simulating the false joint in an experimental animal's shin fracture comprises proximal and distal bases in the form of a sector equal to 3/4 of a ring. Proximal base consists of 2nd and 4th levels, distal base consists of 5th and 7th levels, sectors of levels 2, 4 and 5 are placed one below the other while 7th level sector is flipped through 180° in a horizontal plane in relation to the sectors of levels 2, 4 and 5. Two threaded fixing rods connect the 2nd and 4th, 4th and 5th, 5th and 7th levels, and one fixing rod connects the 2nd level with 4th and 5th levels. Each base comprises two pairs of bolted pin fixators. Model reproducibility makes 100%.

EFFECT: simulating the false joint having the characteristics maximum close to the real clinical process.

2 cl, 8 dwg, 1 ex

FIELD: medicine.

SUBSTANCE: what is presented is a device which comprises two handles with branches at working ends of which provided is a line of teeth with pointed tops and a widened base. The teeth are arranged on a semi-circumference on an anterior working surface and at edges of the branches. When closed, tops of the anterior teeth of the upper branch are arranged in front of the tops of the teeth in the lower branch. Tops of the side teeth of the upper branch project into a lumen between the tops of the side teeth of the lower branch. Each branch between the lines of the teeth comprises a calibrated long window at its greater edge; the last tooth is followed by a notch forming a lumen between tooth-free surfaces of the branches when closed. Preferentially, between the handles there is a plate spring holding the handles separated apart to be brought together until the teeth of the upper and lower branches close. One end of the plate spring is attached to an internal surface of an end of the upper handle, while the other end of the spring is provided on an internal surface of the middle portion of the lower handle and sliding longitudinally thereon.

EFFECT: uniform bite wound simulation in all experimental groups by increasing more accurate localisation, form, size and nature of a multiple wound that increases the quality of comparative analysis of various wound healing techniques.

2 cl, 5 dwg

FIELD: information technology.

SUBSTANCE: method of evaluating correctness of actions of a femoral aortography trainee is realised using a virtual computer simulator, wherein a special-purpose peripheral device for simulating tool actions transmits control signals to a special-purpose software system for controlling a computer operating scene; in said system, a unit for simulating computer operating scene objects based on mathematical models creates a computer operating scene; information on events relating to the computer operating scene is transmitted to a unit for recording events during operation simulation, which records actions of the trainee and/or changes in the state of the operating scene with the required sets of parameters; the unit for recording events during operation simulation transmits information to a unit for real-time evaluation of significance and correctness of performed actions, which processes recorded events by applying thereto formalised criteria for significance and correctness of performed actions; the unit for real-time evaluation of significance and correctness of performed actions transmits the set of estimates of actions of the trainee at the end of operation simulation to a unit for final evaluation of actions.

EFFECT: high effectiveness of evaluating actions of a femoral aortography trainee.

1 dwg

FIELD: medicine.

SUBSTANCE: invention refers to experimental medicine, namely to simulating inflamed paranasal sinuses accompanying chronic tobacco smoking. That is ensured by placing the rats into an inhalation chamber wherein tobacco smoke containing 98 mg/m of carbon monoxide is supplied. The cigarettes containing tar 11 mg and nicotine 0.8 mg are used. The tobacco smoke is supplied at a rate of 1 cigarette per 8-10 minutes for 1.5 hours daily for 6 months.

EFFECT: method provides adequate simulation of the inflammation processes accompanying chronic exposure to tobacco smoke, which enables studying of the pathogenesis and control of the inflammation, using correction methods, including drug therapy, while reducing the cost of providing the model and reducing the labour intensity of the simulation procedure.

2 dwg, 1 tbl,1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to the field of biochemistry, in particular to clostridial neurotoxins with a modified persistence. Claimed is a polypeptide, containing HC-domain, the first and, at least, one additional LC-domain with amino acid sequences, at least, 90% identical to the respective sequences of a neurotoxic component of botulotoxin of a serotype A, B, C1, D, E, F or G. Also claimed are nucleic acid, an expression vector and a host cell, intended for the expression of the said polypeptide. Also claimed are a method of obtaining and application of the said polypeptide, including as a component of a pharmaceutical composition, for treatment of a condition, associated with hyperactive cholinergic innervations of a muscle or a exocrine gland, and for cosmetic procedures, associated with wrinkles.

EFFECT: invention makes it possible to controllably vary a period of activity of clostridial neurotoxins.

12 cl, 1 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: invention can be used to identify a high risk of developing impaired glucose tolerance in patients with stable effort angina with underlying administering beta-adrenergic blocking agents with no additional vasodilating properties. Therapy is preceded by conducting 2 exercise tests on the same day to achieve a threshold load power according to the same protocol, initially and 2 hours after administering a single dose of the beta-adrenergic blocking agents. If observing an interval gain of 120 seconds and more from the beginning of the load to the angina attack and/or reduction of an ischemic ST segment on the electrocardiogram not less than 1 mm at the 2nd load as compared to the 1st load, a risk of impaired glucose tolerance is considered to be high. A glucose tolerance test is carried out in these patients 4-5 weeks after the scheduled administration of the beta-adrenergic blocking agents. If impaired glucose tolerance is detected, administering the beta-adrenergic blocking agents is withdrawn. If the 2nd load as compared to the 1st load shows an interval to the angina attack and/or reduction of the ischemic ST segment on the electrocardiogram at a depth not less than 1 mm increasing less than by 120 seconds, a risk of developing impaired glucose tolerance is considered to be negligible. Treatment of these patients with the beta-adrenergic blocking agents is continued without the glucose tolerance test required.

EFFECT: method provides preventing carbohydrate metabolic disorders by the early identification of the high risk of developing impaired glucose tolerance in the given patients by detecting a compensatory increase of the glucose consumption with insulin resistance and a lower availability of free fatty acids to provide myocardial energy needs.

6 ex

FIELD: medicine.

SUBSTANCE: method involves intravenous allogeneic transplantation of multipotent mesenchymal stromal cells recovered from the placenta in an amount of 6 mln cells/kg. Additionally, haemopoietic stem cells recovered from umbilical blood in an amount of 300 thousand cells/kg are introduced.

EFFECT: method enables reducing the number of cytogenetically changed cells, promotes activating myeloid tissue regeneration in old laboratory animals.

4 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method involves providing an anaesthetic support with using the preparations: midazolam, fentanyl, propofol by bolus injections and infusions, artificial pulmonary ventilation by gas mixture O2:N2O=1:2 inhalations. Postoperative nausea and vomiting are prevented by intravenous administration of ondansetron 8 mg and dexamethasone 8 mg at the stage of induction, and by intravenous administration of clonidine 0.0016±0.0002 mg/kg/h in infusions or 0.02-0.025 mg in bolus doses up to 0.0016±0.0002 mg/kg after intubation of the trachea within the first hour of operation with having blood pressure and heart rate controlled.

EFFECT: method is safe and high-effective for preventing postoperative nausea and vomiting.

1 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely surgery, and may be used for preventing peritoneal adhesions. That is ensured by administering Licopid 0.01 g once a day orally for 6 days from the first postoperative day without regard to the extent of operation.

EFFECT: method enables reducing the postoperative adhesion formation with no side effects and reducing the length of treatment.

FIELD: medicine.

SUBSTANCE: invention refers to medicine and can be used in treating chronic inflammatory diseases accompanied by immune deficiencies. To do this, having a clinical-ambulatory immune deficiency stated, the therapeutic stage of a primary disease is followed by introducing to a patient of one-group fresh umbilical blood in an amount of three millilitres mixed with one gram of a broad-spectrum antibiotic, and one millilitre of a local anaesthetic. The above is injected three times into an infrascapular region every two days.

EFFECT: using the given invention enables providing the manifested and stable effect of increasing values of the humoral component of the immune system enabling preventing manifestations of secondary infections.

10 tbl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: what is presented is an agent for poly(ADP-riboso)polymerase inhibition. The agent represents 7-methylguanine(2-amino-7-methyl-1H-purin-6(7H)-one)- a purine derivative of formula (I) The agent has shown the efficacy higher than that in 7-methyl-xanthine and is non-toxic for the human body.

EFFECT: agent can be used in treating conditions caused by necrotic cell death: stroke, myocardial ischemia, diabetes and complications thereof, shock, neurotrauma, arthritis, colitis, allergic encephalomyelitis and other inflammations.

1 dwg, 2 ex

FIELD: biotechnology.

SUBSTANCE: composition is proposed which comprises stem cells of human amniotic fluid with the phenotype CD73+/CD90+/CD105+/CK19+, nutrient medium, erythropoietin, epidermal growth factor, and collagen taken in an effective amount.

EFFECT: invention enables to increase the proliferative potential and viability of the cells, while simultaneously providing cytoprotective effect on the cells of the transplant and stimulation of migration and proliferation of patient's own cells, and also to reduce significantly the concentration of injectable cells and to activate vascularisation and regeneration at the defect site and can be used in therapy for elimination of congenital and acquired defects of soft tissue arising as the result of injuries, after removal of tumors, congenital diseases, age-related changes or other damages.

2 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: invention represents a pharmacological geroprotective composition, which includes a polyphenol component, vitamins and microelements, humic acids, containing polyphenol components, vitamin C, vitamin A, iron (II) chloride and selenium (IV) dioxide, with the composition components being in a specified ratio in wt %.

EFFECT: increased life expectancy and retardation of tumour development.

2 cl, 3 dwg, 2 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of general formula (I) or pharmaceutically acceptable salts thereof, where Alk is an C1-C6alkyl group; G is C=O and Q is CR51R52 or NR51, where R51 and R52, being identical or different, independently denote H, C1-C6alkyl, optionally substituted with a substitute selected from a group comprising carboxy, phenoxy, benzyloxy, C1-C6alkoxy or hydroxy; C3-C6cycloalkylC1-C6alkyl; phenylC1-C6alkyl, optionally substituted with a halogen; phenylamidoC1-C6alkyl; phenylC1-C6alkylamidoC1-C6alkyl, optionally substituted with a C1-C6alkoxy group; or R51 and R52, together with a carbon atom with which they are bonded form a C=O or C2-C6alkenyl group, optionally substituted with a phenyl; M1 is CR49, where R49 is H; M2 is CR50, where R50 is H; R38 is H, C1-C6alkyl, substituted with a phenoxy group; C3-C6cycloalkylC1-C6alkyl; arylC1-C6alkyl, optionally substituted with 1 or 2 substitutes selected from a group comprising C1-C6alkyl, C1-C6alkoxy, C1-C6alkoxycarbonyl, carboxyl, N-methylamido, hydroxy, C1-C6alkoxyC1-C6alkoxy, C1-C6alkylthio, C1-C6alkylsulphanyl, cyano, halogen, perfluoroC1-C6alkyl, nitro, formyl, hydroxyC1-C6alkyl and amino, wherein the aryl moiety is a phenyl or naphthyl; and heteroarylC1-C6alkyl, where the heteroaryl moiety is pyridinyl, optionally substituted with 1 or 2 groups selected from C1-C6alkoxy or hydroxyC1-C6alkyl, pyrazolyl or isoxazolyl, substitute with 1 or 2 C1-C6alkyl groups; R47 and R48 is C1-C6alkyl. The invention also relates to specific compounds, a method of reducing or weakening bitter taste, a composition of a food/non-food product or beverage or drug for reducing or lightening bitter taste and a method of producing a compound of formula (I).

EFFECT: obtaining novel compounds which are useful as bitter taste inhibitors or taste modulators.

37 cl, 6 dwg, 12 tbl, 186 ex

FIELD: medicine.

SUBSTANCE: method involves intravenous allogeneic transplantation of multipotent mesenchymal stromal cells recovered from the placenta in an amount of 6 mln cells/kg. Additionally, haemopoietic stem cells recovered from umbilical blood in an amount of 300 thousand cells/kg are introduced.

EFFECT: method enables reducing the number of cytogenetically changed cells, promotes activating myeloid tissue regeneration in old laboratory animals.

4 tbl, 1 ex

Up!