A method for the diagnosis of multiple sclerosis
The invention relates to medicine, namely, neurology, and can be used for the diagnosis of multiple sclerosis. Examine the blood plasma by the method of the wedge dehydration. A drop of plasma pre-dried on a glass slide. Then analyze the structure of the resulting facies under the microscope. If the Central zone facies abnormal structures such as the Sierpinski carpet diagnosed with multiple sclerosis. The method is simple and affordable and allows faster diagnosis of multiple sclerosis. 1 Il., 1 PL.
The invention relates to medicine, namely, neurology, and can be used for the diagnosis of multiple sclerosis in patients with clinical manifestations of this disease and for the differential diagnosis in doubtful cases.
Multiple sclerosis (PC) - chronic demyelinizing disease of the Central nervous system affecting young adults and accompanied by the loss of health, and in some cases, in the later stages of the disease, is the ability to move and self-help that makes the social importance of this issue. The prevalence of PC is growing in many of strangleweed [2, 7].
Over the last decade PC from incurable diseases categorized as partially managed. Under these conditions, accurate diagnosis is an essential tool in determining the subsequent treatment. The problem of diagnostics PC is very important. According to the clinic of neurology of the medical University n.a. Acad. I. P. Pavlova 1998-1999 frequency of wrong - false-positive or false-negative diagnosis for referral to hospital was 37% (30 of 81). The majority of patients (66) had an “experience” ill Denmark from 1 to 10 years and held various diagnostic procedures .
Become important methods of early diagnosis and supportive care, which contributes to the longer life expectancy of patients and diminished among them the percentage of heavy persons with disabilities .
The PC diagnosis continues to be based primarily on clinical data . There is not pathognomonic laboratory test, allowing unequivocally establish the diagnosis PC . For a number of diseases, such as acute disseminated encephalomyelitis, neurobrucellosis, neurosarcoidosis, neurospin, chronic fatigue syndrome, vascular and metabolic disorders and many other conditions can iinformative and available research confirming the diagnosis of PC. The main disadvantage of that detected in patients with PC MRI changes are not specific and are found in many States, not related to PC. The most common diseases and conditions which by the nature of the changes in MRI can be similar to the PC include: vascular diseases of the brain (vasculitis, migraine, hypertension, subcortical arteriosclerotic leucoencephalopathy, periventricular leucomalacia), infectious and inflammatory diseases (Lyme disease, neurosarcoidosis, acute disseminated encephalomyelitis, progressive multifocal leucoencephalopathy, encephalitis in HIV infection). The number, size and localization of foci of poorly correlate with clinical symptoms, degree of disability of the patient and give very little information regarding prognosis [5, 7].
Use as a contrast agent gadolinium to determine the activity of the process makes diagnosis unnecessarily expensive, in addition, increased accumulation of contrast in the area of “foci” is sometimes not associated with clinical worsening. The use of MRI for monitoring of the disease or the effectiveness of therapy and “old” is able to significantly shrink in size and even disappear .
Non-invasive, safe and easy investigation of evoked potentials (EP) are usually not sufficiently informative for the diagnosis of PC. Using visual EP can be identified involvement of the optic nerve in patients without symptoms of optic neuritis, but, unfortunately, a fairly large list of conditions that can cause similar changes in the latency and shape of the peaks visual EP .
Use with suspected PC electroencephalography, electroneuromyographic and electromyography is inadequate because informative in Diagnostika PC is very small .
Among the methods of laboratory diagnostics PC is considered to be the most informative study of cerebrospinal fluid (CSF) for the presence of at least two oligoclonal bands of immunoglobulins (CRC). However, this method allows only indirectly to testify in favor of the PC, but does not confirm the diagnosis. The CRC can be detected in CSF and in many other States. The index of diagnostic accuracy in conventional practice does not exceed 65-70%. The presence of AGR in CSF is absolutely not very interesting, if they are detected in serum .
Closest to the proposed invention are methods immunologicheskaja immune system in genetically predisposed persons. To confirm this diagnosis and more information about the pathogenesis of the disease are the study of blood groups, HLA markers, antibodies, cell subpopulations (CD4+, CD8+, and others), the number of b - and T-lymphocytes. However, it does not give specific for PC information, but requires special equipment, a lot of money and time [3, 6].
For the prototype of the present invention, the selected method for the diagnosis of PC, including the study of blood plasma . In this way reveals the breakdown products of myelin. The disadvantage of the prototype method is:
- low specificity, because the myelin is destroyed, and in other pathological processes in the Central nervous system (tumor, vascular lesions, trauma, and others);
- long waiting for the result of the analysis, as used diagnosticum requires the simultaneous study of plasma 80 patients that involves the accumulation and long-term storage of frozen plasma of patients. Have several months to wait for the result, which delays the timely start of adequate treatment.
Interesting method wedge dehydration, which allows to make visible the molecular organization of biological fluids by transferring it to the macro-level . After vysya amount of organic substances from the center to the periphery . Method wedge dehydration was first proposed for the study of urine . The authors of this method was discovered the phenomenon of pathological crystallization of salts in the protein environment with the active formation in the kidneys (the phenomenon of Shatokhina-Shabalin). Subsequently, they have developed methods of diagnosis of Candida urinary tract infections and some diseases of the kidneys on the analysis of patterns of facies urine. The blood plasma was much more complex object of study due to the large variability of its composition and properties. Apart from the determination of zonal structures in the diagnosis are important shape, number and size of cracks, and the identification of pathological structures in the dried film - facies - plasma . Proposed Method of assessing the state of homeostasis”  in which the parameters of zonal structures of the dried drops of biological fluids define homeostasis as normal or pathological, as well as sustainable or unsustainable. But what pathology can be diagnosed, is not specified. And from the point of view of diagnosis and differential diagnosis of PC, this method was not used.
The task of the invention is supposed to increase stultitiam studies of blood plasma is achieved by the plasma from the blood are examined by the method of the wedge dehydration, with a drop of plasma pre-dried on a glass slide and then analyze the structure of the resulting facies under the microscope, and the presence in the Central zone facies abnormal structures of the type field Sierpinski diagnosed with multiple sclerosis.
We examined 165 people with various neurological and oncological diseases (see table) at the age from 20 to 76 years. Of them with demyelination diseases - 40 people, including 34 - PC.
The difficulties of differential diagnosis of PC was observed in 7 cases.
All patients were given a comprehensive clinical-neurological and laboratory research, including the study of neurological status, careful analysis of anamnestic data, the study of biochemical blood parameters and immune status, neuroophthalmology, vibrotest, electroneuromyography, visual evoked potentials, computed tomography of the brain (as indicated). The blood plasma of all patients were studied using the wedge dehydration. Using pipetochnoe of microdontia drop of plasma volume of 0.01 ml was applied on a clean glass slide. This od microscope in transmitted light.
When comparing the results of the clinical examination and facies patterns of blood plasma was found that all patients have a PC in the Central zone facies identified similar education - anomalous field type Sierpinski carpets. The drawing shows a micrograph facies blood plasma of the patient X (a - increase h, B - increase h), arrows indicate anomalous field type field Sierpinski in the Central zone facies.
Patients in controversial cases where the clinical picture of the disease resembled PC, the above sign was not detected. For additional methods of research (neuroophthalmology, visual evoked potentials, CT) allowed to exclude the diagnosis of PC. Patients with other demyelination diseases of the nervous system (OVDP and hwdp) the presence of abnormal fields of type Sierpinski carpets in the Central zone facies have been identified, out of 19 cases of myasthenia they were observed in 5 patients, but has been a single representation. In addition, these patients revealed other pathological patterns that were not in the facies plasma of patients with PC (grebeshkov patterns and the structure of the sheet type in the peripheral zone).
P is octavos Vienna on an empty stomach, centrifuged for 10 minutes at 1500 rpm and the plasma in an amount of 0.01 ml with pipetochnoe of microdontia put on a clean glass slide and dried at room temperature in a closed Cabinet. After 18-24 hours, the resulting facies are examined under a microscope in transmitted light at low magnification. If the Central zone facies anomalous fields such as carpets Sierpinska about make a conclusion about the possibility of a PC.
To illustrate the claimed diagnostic technologies provide examples of the practical application of this method.
Example 1. Patient K., aged 40, case history No. 0307341, was admitted to the clinic of nervous diseases of the Nizhny Novgorod regional clinical hospital. N. A. Semashko routinely 24.03.03. Upon receipt complained of dizziness, staggering when walking, weakness, fatigue, impaired night's sleep, decreased background mood. From the anamnesis it is known that she has been ill since February 2003, when against the background of increasing blood pressure to 150/100 mm RT.article appeared dizziness with nausea and vomiting, gait disturbance, weakness. Despite the stabilization of numbers of blood pressure, the above complaints included the ri study neurological status showed instability in the Romberg position, intentsionnogo jitter when running coordinatory samples, more pronounced in the lower extremities, the recovery of tendon reflexes with the expansion of their zone, the absence of pathological wrist and stop signs, sensitive disorders. The differential diagnosis was carried out between vascular lesions of the brain and demyelinizing disease. Laboratory research has revealed: ASAT - 24 u/l, Lat - 26 u/l, LDH - 208 u/l, alkaline phosphatase - 403 u/l, fibrinogen - 2,75 g/l, the fraction of blood proteins albumin 62%, Alfa - 2%, A2 - 7%, beta - 11%, gamma 18%, total protein 68 g/l, lipid profile - total cholesterol - 4.4 g/l, Gagliardi - 1.4 mmol/l, cholesterol VLDL - 0.6 mmol/l, HDL - 1.3 mmol/l, beta-lactoprotein - 3.4 g/l, haemoglobin rate is 2.4, the CEC 105 IU/ml Deviations from the norm there. The immunological: IgA - 2,71 g/l, IgM - 1,21 g/l, IgG - 23,53 g/l (above the norm).
When examining neuroophthalmology: pathological changes on the main day is not identified, the disks of optic nerves from both sides pale pink color, border, clear, artery and vein of the eye fundus is not changed. Visual evoked potentials - a variant of norm. The vibrotest - within normal limits. Transcranial Doppler ultrasound significant hemodynamic disturbances did not reveal the Central zone radial cracks and multiple anomalous structure type Sierpinski carpets. Thus, using the method of wedge dehydration plasma from a patient diagnosed clinically probable multiple sclerosis, spinal form (Criteria for the diagnosis of PC C. M. Poser et all, 1983). The diagnosis is confirmed by magnetic resonance imaging brain, which found multiple hypodense lesions in periventricular zone, which is characteristic for demyelinating process.
Example 2. Patient X., 42 years old, medical history, No. 0304775, was admitted to the neurological clinic of NOCB 20.01.03 with complaints of gait disturbance, weakness in the legs, left arm, dizziness, a “burning” sensation in the hips, violation of fluency. The patient feels that he is about 7 years old when I first noticed the diplopia. In the treatment of vascular and nootropic drugs these phenomena were stopped. Deterioration of health arose two years later, when there was a disturbance of vision in his right eye, weakness and numbness in the left foot. Was hospitalized in the clinic of the Nizhny Novgorod regional clinical hospital. N. A. Semashko, which was first diagnosed with demyelinating for what it exacerbation in October 2002, which is associated with the second interrupt pregnancy in connection with intrauterine death of the fetus at 30 weeks. A week after caesarean section occurred gait disturbance, impaired smooth speech, weakness in the right hand. Stationary domiciliary treatment was conducted, including pulse therapy corticosteroid drugs. There has been little trend in the reduction of weakness in his right hand, however, there are disturbances of gait and speech disturbance. Objectively: the patient emotionally labile, diffuse muscular hypotonia, horizontal and vertical nystagmus, ataxic gait, ataxia in the Romberg intentsionnogo jitter when running coordinatory samples, more in the left extremities, decreased strength in the lower extremities and the left hand to 4 points, recovery of tendon reflexes sd, zone expanded, pathological ankle and wrist signs, absence of abdominal reflexes.
Laboratory investigations revealed: the General analysis of blood, urine, ECG within normal limits. Blood sugar - 5.0 mmol/l Fraction of blood proteins - albumin 62%, Alfa - 2%, A2 - 6%, beta - 11%, gamma - 19%, total protein - 77 g/L. lipid profile - total cholesterol 3,8, triglycerides - 1,0, hi serum - 0.49 mmol/l, copper - 16 µmol/l LDH - 256 u/l, alkaline phosphatase - 340 u/l, fibrinogen 2.5 g/l, the CEC 15 IU/ml the Immunological: IgA - of 2.38 g/l, IgM - of 5.26 g/l, IgG - 14,63 g/L. Reschinovye test: RCC - 1.2 mg%, NBSC-155 mg%. The fundus of the eye: angiopathy of the retina initial (hypotonic type). The vibrotest found a violation of vibration sensitivity at all frequencies. Visual evoked potentials increased latencies and decreased amplitudes of all selected components of the evoked potentials. These symptoms are typical of the PC.
Structure facies of blood plasma in the peripheral zone of the arcuate cracks and abnormal structure of the Arnold tongues in the Central zone is a dense network of cracks and multiple anomalous structure type Sierpinski carpets.
Clinical diagnosis: Clinically significant multiple sclerosis, spinal form, remitirse type of flow.
Sources of information
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4. Neroev Century Century, Charles I. H., Shapiro, A. L., Samsonova A. M. Psychophysical symptoms of demyelinating lesions of the optic nerve in multiple sclerosis. //Vestnik of ophthalmology, Vol 117, No. 6, 2001. - S. 29-33.
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6. Totolan N. A., Graziela I. C., Klimovich B. B., totolan A. L. Free light chains of immunoglobulins in biological fluids of patients with multiple sclerosis. //Journal of neurology and psychiatry. S. S. Korsakova, Vol 97, No. 5, 1997. - S. 34-39.
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A method for the diagnosis of multiple sclerosis, including the study of blood plasma, wherein the plasma from the blood are examined by the method of the wedge dehydration, with a drop of plasma pre-dried on a glass slide and then analyze the structure of the resulting facies under the microscope and in the presence in the Central zone facies abnormal structures such as the Sierpinski carpet diagnosed with multiple sclerosis.
FIELD: medicine, psychiatry.
SUBSTANCE: one should isolate DNA out of lymphocytes of peripheral venous blood, then due to the method of polymerase chain reaction of DNA synthesis one should amplify the fragments of hSERT locus of serotonin carrier gene and at detecting genotype 12/10 one should predict the risk for the development of hallucino-delirious forms of psychoses of cerebro-atherosclerotic genesis.
EFFECT: more objective prediction of disease development.
FIELD: medicine, urology.
SUBSTANCE: one should conduct subcutaneous prevocational tuberculin test and, additionally, both before the test and 48 h later it is necessary to perform the mapping of prostatic vessels and at decreased values of hemodynamics one should diagnose tuberculosis. The information obtained should be documented due to printing dopplerograms.
EFFECT: more reliable and objective information.
1 ex, 1 tbl
FIELD: molecular biology.
SUBSTANCE: the suggested innovation deals with the fact that nucleic acids should be isolated directly out of the sample without pipetting stage but with the help of interconnected reservoirs being prepared beforehand. The above-mentioned vessels should be applied either separately or being interconnected according to standard microtitrating format. The sample should be mixed with a lyzing buffer and nucleic acids are bound with matrix in closed system including, at least, two interconnected reservoirs. Forced movement of sample's mixture and buffer back and forth from one reservoir into another one for several times through narrow passage provides their thorough intermixing. The method provides quick and safe isolation of nucleic acids.
EFFECT: higher efficiency.
44 cl, 4 dwg, 1 ex
FIELD: medicine, phthisiology, microbiology.
SUBSTANCE: diagnostic material is poured preliminary with chlorohexidine bigluconium solution, homogenized, kept at room temperature for 10-12 h and centrifuged. Precipitate is poured with Shkolnikova's liquid medium, incubated at 37oC for 3 days, supernatant part of Shkolnokova's medium is removed, fresh Shkolnikova's medium is added, and precipitate is stirred and inoculated on the dense cellular egg media. Sensitivity of the strain is determined in 3 weeks by the presence of growth in the control tube only. Invention provides enhancing precision and reducing time for assay. Invention can be used in assay for medicinal sensitivity of tuberculosis mycobacterium.
EFFECT: improved assay method.
FIELD: medicine, biotechnology, pharmacy.
SUBSTANCE: invention relates to agents used for treatment of pathological states associated with disorder of synthesis of neuromediating substances. Method involves the development of pharmaceutical composition and a method for it preparing. Pharmaceutical composition represents subcellular synaptosomal fractions: synaptic membranes, "light" synaptosomes and "heavy" synaptosomes prepared from gray matter of cerebral hemispheres from experimental animals based on the goal-seeking modification of humoral mediators of nerve endings transformed to synaptosomes in development and regression of malignant processes. The composition provides inhibiting the growth of tumor cells, to elevate span-life of patients with ascite Ehrlich's sarcoma, breast adenocarcinoma Ca-755, Wolker's carcinosarcoma-256.
EFFECT: valuable medicinal and anti-tumor properties of composition.
12 cl, 3 tbl, 3 ex
SUBSTANCE: method involves carrying out microscopic examination of blood serum samples taken from femoral vein and cubital vein. Femoral vein sample is taken on injured side. The examination is carried out before and after treatment. The blood serum samples are placed on fat-free glass slide in the amount of 0.01-0.02 ml as drops, dried at 18-30°C for 18-24 h. The set of pathological symptoms becoming larger or not changed after the treatment in comparison to sample taken before treatment, and morphological picture of samples under comparison taken from the cubital vein showing no changes or being changed to worse, the treatment is considered to be effective.
EFFECT: enabled medicamentous treatment evaluation in course of treatment to allow the treatment mode to be changed in due time; avoided surgical intervention (amputation); retained active life-style of aged patients.
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: medicine, urology.
SUBSTANCE: the present innovation deals with differential diagnostics of prostatic cancer and other prostatic diseases at the stage of primary inspection. The method includes the detection of PCA and calculation of probability coefficient for prostatic cancer (PCC) by the following formula: where e - the foundation of natural logarithm (e=2.718…), PCA - the level of total blood PCA in ng/ml, V - patient's age in years. At PCC value being above 0.2 one should diagnose prostatic cancer and to establish final diagnosis one should perform polyfocal prostatic biopsy. The method enables to increase accuracy of diagnostics at decreased number of unjustified prostatic biopsies.
EFFECT: higher efficiency of diagnostics.
FIELD: medicine, biology.
SUBSTANCE: invention relates to nutrient medium used for accumulation of cells for the following cytological and/or immunocytochemical analysis carrying out. Invention relates to medium containing salts NaCl, KCl, anhydrous CaCl2, MgSO4 x 6 H2O, MgCl2 x 6 H2O, Na2HPO4 x 2 H2O, KHPO4, NaHCO3, and also glucose and Henx's solution, 10% albumin solution and polyglucin taken in the ratio 1:1:1. Invention provides enhancing the preservation of cells.
EFFECT: improved an valuable properties of nutrient medium.