Method of intraoperative evaluation of peripheral nerves condition
SUBSTANCE: method consists in production of cryostat sections of nerve, washing them in two portions of maleate buffer, incubation in incubation medium. Incubation is carried out for 30-60 minutes and incubation medium has the following composition: acetylcholine iodide 10 mg; 0.1 M Na-maleate buffer 2 ml; 0.1 M sodium citrate 4 ml; 0.03 M copper sulphate 4 ml; 0.005M potassium ferricianide 4 ml; medium pH constitutes from 7.4 to 7.6 and temperature is from 50 to 55°C.
EFFECT: method by invention makes it possible to evaluate viability of peripheral nerves in short term, which is necessary during reconstructive-restoring and highly technological neurosurgical operations.
The invention relates to medicine, namely to reconstructive microsurgery, and is intended for intraoperative assessment of the state of the peripheral nerves.
Assessment of peripheral nerves, bankirovanii fragments of nerves is of considerable interest to the neurosurgeon, because the outcome of the restorative and reconstructive surgery depends on the viability stitched the nerves. To assess whether the nerve sprouting of nerve fibers, stored in damaged nerve trunk exchange of the mediator, whether axonal current, is impossible without use of special morphological methods.
Known electrophysiological methods of assessing conduction in the nerve: electrical stimulation of the nerve, electromyography (Odinak M. M., Zhivolupova S. A. Diseases and injuries of the peripheral nervous system (generalization of clinical and experimental experience): a guide for physicians. - M.: spec lit, 2009, 384). The pulse caused by electrical stimulation of the nerve, is sent by motor, sensory and mixed nerves, and the characteristics of the pulse are measured by recording potentials from the muscles, either directly from the nerve. If the nerve is damaged and has not happened yet restore the contact with the innervation targets, the value of these methods decreases dramatically. In addition, they cannot be used in �burden restorative and reconstructive microsurgical procedures due to the complexity of determining the projection of the thin branches of the nerves.
The closest technical solution (prototype) of the proposed method is the method of Karnovsky M. J., L. Roots (Karnovsky M. J., L. Roots, 1964 A "direct-coloring" Thiocholine method for cholinesterase, J. Histochem. Cytochem., 12, 219-22), which is used to identify the positive activity of acetylcholinesterase (ache) in nervous tissue, definition nervous system accessories guides and study the characteristics of the postsynaptic membrane in the neuromuscular synapses of somatic muscles (Nikolaev, G. M. Changes of acetylcholinesterase activity in motor endings in striated muscle during hypoxic hypoxia / G. M. Nikolaev // Archives of pathology, - 1970. - No. 3, pp. 61 - 65).
The enzyme acetylcholinesterase is formed in the neuron soma and transported along the axon to the presynaptic nerve endings or neuromuscular synapses (Dixon M., Webb E., Enzymes, TRANS. from English., T. I, M., 1982, pp. 363-364). Positive reaction to ache is a symptom of conservation synthetic and transport processes in soma of the neuron and its processes. A positive reaction can occur only in the axial cylinders of conductors and does not directly depend on the state of the myelin sheaths. Negative reaction to ache suggests that in this fragment of the nerve yet sprouting fibers.
Determination of the activity of acetylcholinesterase (ache) by Karnowski-roots trail runs�explained as follows: pick up the stuff, then make frozen sections, fixed samples in 10% formalin to one day, washed with two portions of sodium malatova buffer for 5 minutes each, incubated in a special environment: acetylcholine iodide (10 mg; 0.1 M Na-malaty buffer 2 ml; 0.1 M sodium citrate 4 ml; 0.03 M copper sulfate (4 ml; 0.005 M potassium ferricyanide 4 ml, an inhibitor of non-specific cholinesterase (1.0 ml 10-4 M ISO-OMPA) at 4°C, pH 6.0 for 24 hours, washed with distilled water, put in conditioner mikroskopiruût. The total duration of the reaction is 18-24 hours.
This method is specific for detection of a certain faction acetylcholinesterase.
The disadvantages of this method are as follows: the complexity and duration of the procedure, resulting in the inability to use it intraoperatively.
The proposed method is to simplify procedures and reduce the time for determining the state of the peripheral nerve.
This object is achieved in that changes the mode of incubation: the incubation time from 30 to 60 min, and the composition of the incubation medium: no nonspecific inhibitor of cholinesterase, pH ranges from 7.4 to 7.6, temperature between 50 to 55°C.
The proposed method for the assessment of peripheral nerve ache activity allows for a short period of time to conduct a study of their viability, which gives in�possibility to use it intraoperatively.
The novelty of the proposed method lies in the fact that significantly reduced the incubation time, the incubation medium has a simpler composition: acetylcholine iodide (10 mg; 0.1 M Na-malaty buffer 2 ml; 0.1 M sodium citrate 4 ml; 0.03 M copper sulfate (4 ml; 0.005 M potassium ferricyanide 4 ml, and its pH ranges from 7.4 to 7.6, a temperature of 50 to 55°C.
The technical solutions that have the signs consistent with the distinctive features of our proposed method, not identified that allows to make a conclusion about conformity of the proposed method the criterion of "inventive step".
The proposed method is as follows: during the operation, take a piece of nerve, then make frozen sections. The resulting paste slices on glass slides, washed in two portions malatova buffer, are placed in the incubation medium of the following composition: acetylcholine iodide (10 mg; 0.1 M Na-malaty buffer 2 ml; 0.1 M sodium citrate 4 ml; 0.03 M copper sulfate (4 ml; 0.005 M potassium ferricyanide 4 ml, at a temperature of 50 to 55°C, pH 7.4 and 7.6. The incubation time of 30-60 minutes. After incubation spend microscopy of slices for the detection of acetylcholinesterase activity (ache). If the response to acetylcholinesterase positive, the nerve is considered to be viable; if negative - the nerve is assessed as non-viable. Sections were dehydrated, sign in balm, photographed DL� documentation.
Example 1. Patient T. 27 years old, diagnosis: partial left-sided paralysis of the facial nerve. During the operation to determine the status of the facial nerve was taken cut of the nerve length 0.3 cm Segment of the nerve was taken to the laboratory, prepared frozen sections of 40 µm, washed them in two portions malatova buffer, were placed in incubation medium with a pH of 7.4, a temperature of 52°C, incubation time 45 min. After 45 minutes, the sections were washed in distilled water and made microscope. As a result, it was found that the activity of acetylcholinesterase in the barrel is positive, the trunk of the nerve is viable. Then there began the restoration of the damaged facial nerve using a nerve graft is sewn between the trunk of the facial nerve and its zygomatic branch. As nerve autograft was used sural nerve. Achieved restoration of facial expressions left side of his face.
Example 2. Patient M., 35 years was admitted to the hospital with a diagnosis of injury of the right brachial plexus. The operation was performed to restore the integrity of damaged nerves. During surgery it was discovered damage to the median nerve and neuroma throughout. Removed part of the nerve, which contained a neuroma. After the removal of the damaged nerve arose the need for grafting nerve sural nerve autotr�spuntata, since there was a defect nerve 7 see, then the question arose: not damaged nerve proximal to? To assess the status of the proximal part of the median nerve was taken as part of the nerve 0.3 cm for intraoperative histological assessment of the state. This sample was delivered to the laboratory, where samples of the nerve is frozen in the cryostat and frozen sections were made, 40 μm. The sections were washed in two portions of buffer solution (0.1 M Na-malaty buffer, pH=7,6). Subsequently, the slices were placed in incubation medium with pH of 7.6. Incubated in a thermostat at 55°C for 30 minutes. After 30 minutes, the sections were washed in distilled water and made microscope. As a result, it was found that the nerve at this level is positive acetylcholinesterase activity that gave the opportunity to reduce the distance between damaged nerves and thereby reduce the size of the sural nerve autografts to replace the defect of the nerve. As a result of operative intervention there is mobility of my right hand.
The proposed method was used in the surgical clinic during operations autologous peripheral nerves.
Method of intraoperative assessment of the state of the peripheral nerves by identifying positive acetylcholinesterase activity, which is manufacturing cryostat�x slices of the nerve, washing them in two portions malatova buffer, incubation in the incubation medium, characterized in that incubation is carried out in 30-60 minutes in the incubation medium, which has the following composition: acetylcholine iodide (10 mg; 0.1 M Na-malaty buffer 2 ml; 0.1 M sodium citrate 4 ml; 0.03 M copper sulfate (4 ml; 0.005 M potassium ferricyanide 4 ml, pH from 7.4 to 7.6 and a temperature of 50 to 55°C.
SUBSTANCE: functional activity of neutrophilic granulocytes is studied by means of hemiluminiscent analysis, calculated is an index of formation of active forms of oxygen (IFAFO), representing the ratio of the area under the curve of luminol-dependent hemiluminiscence of the neutrophilic granulocytes, induced by a live bacterial suspension of staphylococcus aureus, to the time of achieving the maximum of luminol-dependent hemiluminiscence of the neutrophilic granulocytes, induced by the live bacterial suspension of staphylococcus aureus. If IFAFO value is higher than 100 o.u./s, sensitisation to staphylococcus aureus in patients with allergic rhinitis is diagnosed.
EFFECT: application of the claimed method makes it possible to diagnose staphylococcal allergy in case of allergic rhinitis before beginning therapy and contributes to the optimal selection of the patients' treatment.
1 tbl, 2 ex
SUBSTANCE: invention represents a method for determining a probability of preserving the myocardium following infarction by creating an admission examination-based data array of 7 peripheral blood parameters, 11 biochemical analysis parameters and 6 parameters of standard 12-lead electrocardiogram in 200 patients with the Q-myocardial infarction and 200 patients without the myocardial infarction. The parameters are stratified with respect to 7 intervals, wherein values related to the probability of preserving the myocardium following infarction are derived by calculating a ratio of the patients without myocardial infarction to all the patients with acute coronary syndrome. The probability is evaluated in a specific patient by analysing the above parameters, searching the respective intervals and values related to the probability of preserving the myocardium in the data array. Summing up the derived values enables calculating an integrated index, which is normalised, and a dimension from 0 to 100% is reduced.
EFFECT: invention enables increasing the prediction accuracy of preserving the myocardium in the patients with acute coronary syndrome.
1 tbl, 2 ex
SUBSTANCE: group of inventions refers to medicine, namely to immunology, and can be used in laboratory diagnostics, as a test system and a method for measuring interferon alpha (IFN-α) antiviral activity in human blood serum. The test system for measuring IFN-α activity in the human blood serum comprises diploid cells, a virus-containing fluid and standard human interferon-α (IFN-α). As the diploid cells, the test system comprises cells of the characterised line of the diploid cells - human M-20 fibroblasts at 20-33 passages cultured in the medium with added 10% fibrinolytically active plasma (FAP). As the virus, the system contains the A vesicular stomatitis virus (VSV) adapted to M-20 cells, the Indiana strain; the test system additionally contains a viral dye based on two fluorochromes - acriflavine and rhodamine C. The group of inventions also involves a method for measuring IFN-α activity in the human blood serum with the use of the developed system.
EFFECT: using the given inventions enables the quantitative good-reproducible measurement of IFN-α activity in the examined blood serum sampled by the luminescent microscopy of the vital preparations coloured in a fluorochrome-based vital dye.
3 cl, 4 dwg, 1 tbl, 4 ex
SUBSTANCE: invention describes a diagnostic technique for infected pancreonecrosis with establishing the indications for a surgical intervention by examining a patient, wherein acetic, propionic, butyric and isovaleric acids are measured in the blood by gas chromatography, and if the acetic acid concentration is more than 0.11 mmole/l enables stating infected pancreonecrosis, and if the concentration of any of three acids: propionic more than 0.095 mmole/l, butyric more than 0.0035 mmole/l, isovaleric acid more than 0.0003 mmole/l enables stating infected pancreonecrosis with active development of an anaerobic infection requiring one of versions of the surgical intervention.
EFFECT: invention enables providing the objective and accurate diagnosing of pancreonecrosis transformation into the stage of infection by using the qualitative parameters with no undesired side effects.
SUBSTANCE: invention relates to medicine, namely to a method of predicting a probability of reduction of the glomerular filtration rate (GFR) after 3 months of observation after aortocoronary bypass surgery without artificial blood circulation (ACBS without ABC). The essence of the method consists in the fact that the concentration of a kidney injury molecule of type 1 (KIM-1) is determined in blood serum, the ratio of the biomarker KIM-1 concentrations in two time points after 48 hours and 7 days after the operation is calculated and if its value is higher than 1.5, a conclusion about the probability of (GFR) reduction in the remote period after ACBS without ABC is made.
EFFECT: application of the claimed method makes it possible to predict the probability of the glomerular filtration rate (GFR) reduction after 3 months of observation after aortocoronary bypass surgery without artificial blood circulation in an efficient and accurate way.
1 tbl, 1 dwg, 1 ex
SUBSTANCE: method includes determination of a titre of antibodies to cytomegalovirus, content of 2,3 DPG (2,3-diphosphoglycerate), and oxyhaemoglobin in erythrocytes. If the titre of the antibodies to cytomegalovirus increases to 1:1600, DPG increases to 6.7±0.3 mcmole/ml, content of HbO2 is 95.0±1.7%, with the reduction of the specific optical density of haemoglobin to 0.70±0.01, it is concluded that a threat of anaemia development is formed.
EFFECT: method makes it possible to study the character of haemoglobin oxygenation impairment by means of determination of the specific optical density.
SUBSTANCE: neurovisualisation examination of brain is carried out, Cirs comorbidity index and Kaplan-Feinstein comorbidity index are determined, cochleovestibular syndrome, eye-moving impairments, type of diabetes mellitus are identified. Value of discriminate function (D) is calculated. If D value is higher than zero, diagnosed are consequences of ischemic brain stroke (IBS) with hyperhomocysteinemia (HH), if D is lower than zero, consequences of IBS without HH are diagnosed.
EFFECT: method makes it possible to increase reliability of diagnostics of IBS consequences, which is achieved due to complex analysis of said parameters.
FIELD: veterinary science.
SUBSTANCE: method includes detection of leukocytes, protein, haemoglobin in faeces, and PH-reaction of faeces. A sample of faeces is dissolved in distilled water and applied drop by drop onto appropriate test fields of a test strip designed for urine testing. By variation of colour within 1 minute the reaction is assumed as positive. If pH is less than 7.0 or more than 7.5, and faeces contain soluble protein, haemoglobin and leukocytes at the same time, availability of the inflammatory process in the intestines is confirmed.
EFFECT: invention makes it possible to quickly and accurately diagnose inflammatory process in intestines.
2 cl, 2 ex
SUBSTANCE: invention refers to medicine and aims at diagnosing vaginitis in females of a reproductive age. Expression levels of mRNA of TNF, IL18, GATA3 and TLR4 genes are measured in vaginal smears. Expression levels of mRNA of TLR4/GATA3 and TNF/IL18 genes are related, and the derived threshold cycles are used to predict a probability of vaginitis by formula. If P≤57% for nonpregnant females or P≤68.5% for pregnant females, the absence of vaginitis is stated. If P≥57% for nonpregnant females or P≥68.5% for pregnant females, vaginitis is diagnosed.
EFFECT: invention provides the effective diagnosis of vaginitis in the females of a reproductive age.
3 tbl, 5 ex
SUBSTANCE: group of inventions refers to medicine, namely to a method of analysing ex-vivo if a patient suffering a cancer is expected to respond therapeutically to a method of treating. To this purpose, a biological sample specified in a group consisting of total blood, plasma or serum samples is taken from the patient. The levels IL10 and IFNγ are measured in the above biological sample; the levels are determined with using the respective IL10 and INFγ specific antibodies. The IL10/IFNγ ratio is derived and compared to a threshold. If the IL10/IFNγ ratio of less than 4 shows that the patient is expected to develop the preventive or therapeutic response to the immunogenic composition. The group of inventions also refers to using IL10 and INFγ as biomarkers and a kit for analysing the above method.
EFFECT: using the given method enables predicting the sensitivity of the patient suffering cancer to the therapeutic treatment, as well as obtaining an algorithm for treatment modification for improving the patient's response to the treatment.
12 cl, 2 dwg, 1 ex
FIELD: medicine, hepatology.
SUBSTANCE: one should detect the level of hepato-specific enzymes (HSE) in blood plasma, such as: urokinase (UK), histidase (HIS), fructose-1-phosphataldolase (F-1-P), serine dehydratase (L-SD), threonine dehydratase (L-TD) and products of lipid peroxidation (LP), such as: dienic conjugates (DC), malonic dialdehyde (MDA). Moreover, one should detect the state of inspecific immunity parameters, such as: immunoregulatory index (IRI) as the ratio of T-helpers and T-suppressors, circulating immune complexes (CIC). Additionally, one should evaluate the state of regional circulation by applying rheohepatography (RHG), the system of microhemocirculation with the help of conjunctival biomicroscopy (CB) to detect intravascular index (II). In case of increased UK, HIS levels up to 0.5 mcM/ml/h, F-1-P, L-SD, L-Td, LP products, CIC by 1.5 times, higher IRI up to 2 at the norm being 1.0-1.5, altered values of regional circulation, increased II up to 2 points at the norm being 1 point, not more one should diagnose light degree of process flow. At increased level of UK, HIS up to 0.75 mcM/ml/h, F-1-P, L-SD, L-TD, LP products, CIC by 1.5-2 times, increased IRI up to 2.5, altered values of regional circulation, increased II up to 3-4 points one should diagnose average degree of process flow. At increased level of UK, HIS being above 0.75 mcM/ml/h, F-1-P, L-SD, L-TD, LP products, CIC by 2 and more times, increased IRI being above 2.5, altered values of regional circulation, increased II up to 5 points and more one should diagnose severe degree of process flow.
EFFECT: higher accuracy of diagnostics.
FIELD: medicine, infectology, hepatology.
SUBSTANCE: in hepatic bioptate one should detect products of lipid peroxidation (LP), such as: dienic conjugates (DC), activity of antioxidant enzymes, such as: catalase (CAT)and superoxide dismutase (SOD). One should calculate by the following formula: C = DC/(SOD x CAT)x100, where DC - the content of dienic conjugates, SOD - activity of superoxide dismutase, CAT - activity of catalase. At coefficient (C) values being above 65 one should predict high possibility for appearance of cirrhosis, at 46-645 - moderate possibility and at 14-45 -low possibility for appearance of cirrhosis.
EFFECT: higher accuracy of prediction.
FIELD: medicine, clinical toxicology.
SUBSTANCE: at patient's hospitalization one should gather the data of clinical and laboratory values: on the type of chemical substance, patient's age, data of clinical survey and laboratory values: body temperature, the presence or absence of dysphonia, oliguria being below 30 ml/h, hemoglobinuria, erythrocytic hemolysis, exotoxic shock, glucose level in blood, fibrinogen and creatinine concentration in blood serum, general bilirubin, prothrombin index (PTI), Ph-plasma, the state of blood clotting system. The state of every sign should be evaluated in points to be then summed up and at exceeding the sum of points being above "+20" one should predict unfavorable result. At the sum of "-13" prediction should be stated upon as favorable and at "-13" up to "+20" - prediction is considered to be doubtful.
EFFECT: higher accuracy of prediction.
2 ex, 3 tbl
FIELD: medicine, juvenile clinical nephrology.
SUBSTANCE: disease duration in case of obstructive pyelonephritis should be detected by two ways: either by detecting the value of NADPH-diaphorase activity, as the marker of nitroxide synthase activity in different renal department and comparing it to established norm, or by detecting clinico-laboratory values, such as: hemoglobin, leukocytes, eosinophils, urea, beta-lipoproteides, lymphocytes, neutrophils, the level of glomerular filtration, that of canalicular reabsorption, urinary specific weight, daily excretion of oxalates, arterial pressure, and estimating their deviation against average statistical values by taking into account a child's age.
EFFECT: higher efficiency of detection.
7 dwg, 1 ex, 6 tbl
FIELD: clinical medicine, pulmonology.
SUBSTANCE: one should carry out complex estimation of interleukin-1β) concentration in blood, saliva, bronchoalveolar liquid. Moreover, one should detect distribution coefficient (DC) for IL-1β as the ratio of IL-1β blood content to IL-1β salivary content. At increased IL-1β blood content by 10 times and more, by 2 times in saliva, unchanged level of bronchoalveolar IL-1β, at DC for IL-1β being above 1.0 one should predict bronchial obstruction. The method enables to conduct diagnostics of the above-mentioned disease at its earlier stages.
EFFECT: higher efficiency of prediction.
FIELD: medicine, diagnostics.
SUBSTANCE: the present innovation deals with genetic trials, with diagnostic field of oncological diseases due to analyzing DNA by altered status of gene methylation that take part in intracellular regulation of division, differentiating, apoptosis and detoxication processes. One should measure the status of methylation in three genes: p16, E-cadherine and GSTP1 in any human biological samples taken out of blood plasma, urine, lymph nodes, tumor tissue, inter-tissue liquid, ascitic liquid, blood cells and buccal epithelium and other; one should analyze DNA in which modified genes of tumor origin or their components are present that contain defective genes, moreover, analysis should be performed due to extracting and purifying DNA out of biological samples followed by bisulfite treatment of this DNA for modifying unprotected cytosine foundations at keeping 5-methyl cytosine being a protected cytosine foundation followed by PCR assay of bisulfite-treated and bisulfite-untreated genes under investigation and at detecting alterations obtained according to electrophoretic result of PCR amplificates, due to detecting the difference in the number and electrophoretic mobility of corresponding fractions at comparing with control methylated and unmethylated samples containing normal and hypermethylated forms of genes one should diagnose oncological diseases. The method provides higher reliability in detecting tumors, detection of remained tumor cells after operation.
EFFECT: higher efficiency of therapy.
1 cl, 3 dwg, 4 ex
FIELD: medicine, gastroenterology.
SUBSTANCE: one should carry out diagnostic studying, moreover, on the 5th -6th d against the onset of exacerbation in case of gastric and duodenal ulcerous disease one should detect the content serotonin, histamine and acetylcholine in blood, then during 2-3 wk one should conduct medicinal therapy to detect serotonin, histamine and acetylcholine level in blood again and at serotonin content being by 2-3 times above the norm, histamine - by 1.15-1.4 times above the norm and acetylcholine - by 20-45% being below the norm one should predict the flow of gastric and duodenal ulcerous disease as a non-scarring ulcer.
EFFECT: higher accuracy of prediction.
SUBSTANCE: method involves taking blood from ulnar vein (systemic blood circulation) and from large vein of the injured extremity proximal with respect to lesion focus (regional blood circulation). Spontaneous NST-test value is determined and difference is calculated in systemic and regional blood circulation as regional-to-systemic difference. The difference value is used for predicting clinical course of pyo-inflammatory disease in extremities.
EFFECT: high accuracy of diagnosis.
4 cl, 2 tbl
FIELD: medicine, gastroenterology.
SUBSTANCE: one should introduce biologically active substance, moreover, in patient's blood serum one should detect the content of acetyl choline and choline esterase activity followed by 2-h-long intragastric pH-metry at loading with biologically active substance as warm 40-45%-honey water solution at 35-40 C, and at increased content of acetyl choline being above 1.0 mM/l, choline esterase being above 0.5 mM/l/30 min and pH level being 6.0-6.9 it is possible to consider apitherapy to be useful for treating ulcerous duodenal disease.
EFFECT: higher efficiency and accuracy of detection.
FIELD: medicine, gastroenterology.
SUBSTANCE: it has been suggested a new method to detect pharmacological sensitivity to preparations as acidosuppressors. After the intake of the preparation a patient should undergo fibrogastroduodenoscopy 3 h later, then, through endoscopic catheter one should introduce 0.3%-Congo red solution intragastrically and the test is considered to be positive at keeping red color that indicates good sensitivity to the given preparation, and in case of dark-blue or black color the test is considered to be negative that indicates resistance to this preparation. The suggested innovation widens the number of diagnostic techniques of mentioned indication.
EFFECT: higher efficiency of diagnostics.