Method for evaluating organism reactivity in treating for hepatobiliary system abnormality

FIELD: medicine.

SUBSTANCE: method involves determining separated glycogen content in adportal hepatocytes adjacent to the central vein and taking intermediate position between said zones. Reactivity index is calculated as IR=A/B, where A is the hepatocytes percent having substrate in the amount exceeding 75% from cytoplasm area adjacent to the central vein and hepatocytes occupying intermediate position between the zones, B is the hepatocytes percent having substrate in the amount exceeding 75% from cytoplasm area adjacent to portal tract. The RI value being equal to or less than 0.37, stress-training response is to be determined. RI value being greater than 0.7, activation response is determined.

EFFECT: high accuracy of the method.

1 tbl

 

The invention relates to medicine and can be used to assess the reactivity of the organism in the treatment of diseases of the hepatobiliary system.

Known methods for evaluating the reactivity of the organism. Among the commonly used include test reactivity, the principle of which consists in a comprehensive assessment of the basic vital parameters (heart rate, respiration and other) before and after physical activity with subsequent identification of the relationship with the degree of developing hypoxia (1). Often used methods of assessment reactivity, based on the analysis of the main laboratory and clinical parameters of the patient (2, 3). However, most methods for assessing the reactivity is based on a subjective analysis of views about a particular link adaptive and compensatory mechanisms of the body. Often to assess the reactivity of the organism are bulky, complex calculations (scheme), sometimes requiring additional costly methods of research that is possible only in specialized institutions.

Closest to the proposed method is a method of evaluating the reactivity of the organism, based on qualitative and quantitative analysis of glycogen in liver cells (4). Despite sufficient information content in the above-mentioned method, there is no separate zonal features ritika hepatocytes with glycogen content, that is a significant issue. According to literature data the zonal heterogeneity of hepatocytes is dynamic functional and morphological structure as in norm and in pathology (5). The hepatocytes surrounding the portal tract, belong to the hepatic cells of the 1st zone, adjacent to Central Vienna, belong to the hepatic cells of the 3rd zone, occupying an intermediate position between the 1st and the 3rd, to 2nd zone. This circumstance is due to the different cell zones fundamentally different functions, which will undoubtedly affect the nature of the distribution of glycogen in the cytoplasm of liver cells. Thus, to assess the level of reactivity is set as the percentage of hepatocytes with different content of glycogen, and the uniformity of its distribution in cells. Thus, the high content of glycogen in liver cells and a uniform distribution indicate a high functional activity without any tension elements and correspond to the reactions of increased activation of high levels of reactivity, moderately high with a uniform distribution corresponds to the reaction peaceful activation of high levels of reactivity, moderately high with an uneven distribution corresponds to the calm reaction activation low level, the mind is i.i.d. content with a uniform distribution corresponds to the reaction exercise high levels moderate content with an uneven distribution corresponds reactions workout low levels, etc. (5). The reduced glycogen content is a sign of lower adaptive capacity and apparent dysregulation of reactive adaptive reactions (4). The change of reactivity is often accompanied by the development of a number of immunodeficiency, provokes a complex giperergicakie and adverse reactions. In this regard, there is the feasibility in the verification of the positive impact of the treatment on the reactivity of the organism of patients with chronic opisthorchiasis, which is considered to be prognostically favorable objective sign.

In clinical practice with the aim of deworming today are widely used as herbal drugs - acoral (6), and drug-free tools (the use of recycled products-applicators capable of forming the information-wave analogue of the original drug with inclusion frequency EHF). In experiments with models of toxic hepatitis proved the positive influence of electromagnetic radiation of millimeter-wave-band on morphological and functional state of the liver (7, 8).

The problem posed by the invention, is to improve the accuracy and expand the Arsenal of methods of assessment, re is aktivnosti body.

The task to solve a new way to assess the reactivity of the organism, which consists in determining the quantitative content of glycogen in hepatocytes, and define separate the content of glycogen in the hepatocytes of the first, second and third zones and calculate the reactivity index IL by the formula:

where A - % of hepatocytes with high glycogen levels 2 and 3 zones,

In - % of hepatocytes with high glycogen levels 1 zone, and when the value of R & d equal to 0.37 and below, determine the response of the stress-workout, and a value of IL more than 0.7 determine the reaction activation.

The method was carried out as follows.

In experimental animals after treatment of diseases of the hepatobiliary system produce fence fragments of the liver and its morphological study. Fragments of liver immediately after collection were recorded in 12% neutral formalin. The drugs were prepared by the standard method, was poured into paraffin. Slices with a thickness of 5-6 microns were stained CHIC-reaction on the Mac Manus (9).

Estimated quantity of glycogen in hepatocytes 1, 2, and 3 zones. Using the magic wand tool in PhotoShop counts the content of glycogen in the hepatocytes containing substrate more than 75% of the area of the cytoplasm was taken by us as liver cells with a high content of glycogen is a (corresponds to a reaction of increased activation), from 50 to 75% with moderate (corresponding response training), less than 50% - low (corresponds to the response of stress). For the unification of research within each micropreparative the evaluation was conducted in ten fields of each of the third field of view. Five groups of fragments of the liver, including 2 control, were "blind" study. Morphological evaluation was carried out on the installation, including light microscope "Zeiss Jena", the digital camera Epson-200 using PhotoShop to 6.0. The image of the visual field were entered into the computer using a video camera. Finite linear and optical zoom after the image transfer from the camcorder to the computer was - ×800, 1600. Determined the percentage of hepatocytes with high glycogen levels 1, 2, 3 zones and calculated the index IL by the formula:

where A - % of hepatocytes with high glycogen levels 2 and 3 zones,

In - % of hepatocytes with high glycogen levels 1 zone, and when the value of R & d equal to 0.37 and below, was determined by reaction of the stress of training, and a value of IL more than 0.7 was determined response activation.

The proposed criteria are selected on the basis of the analysis of experimental studies, for which was taken 5 groups of observations. Conducted a comprehensive comparative morphological analysis of liver tissue of Golden hamsters (Crecetus adus), infected with opisthorchiasis. Infection was carried out by feeding the hamsters within 7 days fish - Yelets (Leuciscus idus), caught in the river Tom and examined microscopically for the presence of metacercariae. After controlling the opening two hamsters 30-35 days after infection detected 90 and 95 Marit (proven infection).

All infected hamsters, depending on the received treatment, were divided into 5 groups. In the experiment used 20 pieces of liver from each group.

We have reviewed the biopsies of the two control groups. The control group No. 1 was represented by fragments of the liver healthy (non-infected, sterile laboratory animals) hamsters. The control group No. 2 - the liver of hamsters infected with opisthorchiasis, but not treated (fence material at 35-40 days after infection). The experimental group (main): group # 3 - the liver tissue after treatment with corsola (1.0 ml of 1% aqueous solution introduced through the metal probe 3 times a day for 7 days), group # 4 - the liver tissue after treatment non-medical means - information-wave analogue of opisthorchis ("On") and group # 5 - the liver tissue after treatment of the information-wave analogue of corsola ("e"). Drug-free remedies for deworming prepared in accordance with the methodology (7) and used for the treatment of infected hamsters, and placing the (information-wave analogues) in a container of drinking water for 7 days.

Fragments of liver immediately after collection were recorded in 12% neutral formalin. The drugs were prepared by the standard method, was poured into paraffin. Slices with a thickness of 5-6 microns were stained CHIC-reaction on the Mac Manus (9).

Estimated quantity of glycogen in hepatocytes 1, 2, and 3 zones. Using the magic wand tool in PhotoShop counts the content of glycogen in the hepatocytes containing substrate more than 75% of the area of the cytoplasm was taken by us as liver cells with a high content of glycogen (corresponds to reaction of increased activation), from 50 to 75% with moderate (corresponding response training), less than 50% - low (corresponds to the response of stress). For the unification of research within each micropreparative the evaluation was conducted in ten fields of each of the third field of view. Five groups of fragments of the liver, including 2 control, were "blind" study. Morphological evaluation was carried out on the installation, including light microscope "Zeiss Jena", the digital camera Epson-200 using PhotoShop to 6.0. The image of the visual field were entered into the computer using a video camera. Finite linear and optical zoom after the image transfer from the camcorder to the computer was - ×800, 1600.

On the basis of the analysis of the conducted research was the following conclusions.

Therapy with the use of anthelminthic drug of plant origin corsola, information-wave analogues of opisthorchis and corsola revealed the following (see table 1).

Depending on the type of treatment of chronic opisthorchiasis percentage of hepatocytes first zone (C) with a high content of glycogen was not significantly different between the compared 3, 4 and 5 groups, as well as the content of glycogen (a) in 2, 3 zones (p<0,05). Although the studied parameter in the 2, 3 zones in all the studied groups was actually below.

The reactivity index according to the results view drugs liver healthy hamsters (control group No. 1) amounted to 0.72, while infected with opisthorchiasis, but not treated (control group No. 2) hamsters is 0.37, which is significantly different (p<0.005 percent) and demonstrates the negative impact of opisthorchosis invasion at the level of reactivity of the final master. In all major groups (No. 3, 4, 5) after treatment with corsola, information-wave analogue of opisthorchis and information-wave analogue of corsola the reactivity index amounted to 0.75; 0,73; to 0.74, respectively. A measure of reactivity in the treated groups did not differ from the index of reactivity healthy hamsters (p>0,05), suggesting that therapy suggested ways favorably affect the status is of normal physiological reactivity of the organism as a whole.

No significant differences between the indices of reactivity index, proposed by us, in groups of healthy and treated hamsters leads to the conclusion that antiparasitic (anti opisthorchosis) treatment, including non-medical means (information-wave analogues), has a beneficial effect on the reactivity indices of the body of the final host, which contributes to maintaining full functional state of the liver and prevents the overvoltage. This is especially important as the preservation of the functional state of the liver depends on the preservation of its functions, including detoxifying.

The proposed index of reactivity level of glycogen in liver cells is almost identical to the "reactivity index" of Christ-Quaking, which is calculated by the ratio of lymphocytes to the segmented neutrophills in the blood (<0.3 to stress reaction; 0,31-0,5 - response training; 0,51-0.7 to calm reaction activation; >0,7 - reaction increased activation), which probably suggests a high degree of correlation between these indices.

Thus the proposed method allows to evaluate the reactivity of the organism before and after treatment. In the course of the study we established that therapy chronic is on opisthorchiasis using the information-wave analogue of opisthorchis, information-wave analogue of corsola and plant anti opisthorchosis drug corsola accompanied by a significantly positive indicator of reactivity, indicating a favorable effect of treatment on morphological and functional state of the liver, and the reactivity of the organism as a whole.

Thus, the obtained results allow us to judge the impact of therapy, including non-medical (information-wave equivalent)on the reactivity of the body, moving the body from a state of stress and exercise in a heightened state of activation, which corresponds to the physiological norm with high functional activity without any elements of tension. Identified during the conducted research correlational relationship between reactivity indices proposed "reactivity index", calculated as the ratio of the level of glycogen in the liver cells of the different zones, and the reactivity index", calculated as the ratio of lymphocytes to the segmented neutrophils (Garkavi-Quaking), allows you to focus clinicians on the use of known and available "reactivity index" of Christ-Quaking to assess not only the reactivity of the organism as a whole, but also for predicting the effectiveness of treatment is the actions in the pathology of hepatobiliary system in General and in the treatment of opisthorchiasis in particular.

Table No. 1.

The content of glycogen in hepatocytes 1 and 2, 3 zones in the studied groups (in %, M±m)
groupHepatocytes 1 zone with a high content of glycogenHepatocytes, 2, 3 zones with a high content of glycogen"reactivity index" 2, 3 zones / 1 area
group 1 (healthy)89,9±965,0±6,70,72
group 2 (infected, untreated)40,0±5,3*15±6,9*0,37
group 3 (treated with Corsola)77,0±8,5*58,0±6,0*0,75
group 4 (treated with "About")75,0±8,1*55,0±5,9*0,73
group 5 (treated with "e")71,0±7,9*53,0±6,3*0,74
Note: * - at p<0,05; if no R is the result unreliable.

Comparison group: 2 group 1 group 3, 4 and 5 groups were treated with 2 group (untreated).

References

1. Assist, Lpowell. "Fundamentals of General pathology". ELSBY-SPB, 2002, 618 S.

2. Phharmacy, Ebouaney, Maunaloa. The role of adaptive responses in pathological processes the simple criteria of these reactions in humans. // Regulation of energy metabolism and the body's resistance. Pushchino, 1975, p.172-190.

3. Phharmacy, Ebouaney, Maunaloa, Pembaharuan and other Increasing the body's resistance using adaptive reactions of the training and activation at different levels of reactivity (activation treatment). Methodical recommendations. Rostov n/D, 1983, 34 S.

4. Phharmacy, Avichina, Tschulena. Anti-stress response and activation therapy. M., "IMEDIS", 1998, 656 C.

5. Aselinos, Lieuvin. "Clinical morphology of the liver". M, "Medicine", 1985, 240 S.

6. Masselin M.E., Katukov A.I. of Acoral. Tomsk, LLC "HART", 2001, 20 S.

7. The use of therapy background resonance radiation for relieving pain when neurointelligence diseases. Methodical recommendations. M., 1994, 24 S.

8. Rudin M.V., Ryzhov A.I. Belousov E.V., Kozhemyakin A. M. a Method for preventing liver damage caused by ether anesthesia in the experiment. Patent No. 2153904, Moscow, 2000.

9. Apers "Histochemistry". Publishing house of foreign literature. M., 1962, 964 S.

A method of evaluating the reactivity of the organism in the treatment of diseases of the hepatobiliary system in the experiment, which consists in determining the quantitative content of glycogen in hepatocytes, wherein define separate the content of glycogen in the hepatocytes surrounding the portal is th path, adjacent to Central Vienna and occupy an intermediate position between these zones, and calculate the reactivity index IL by the formula

where A - % of hepatocytes containing substrate more than 75% of the area of the cytoplasm adjacent to Central Vienna, and hepatocytes, which occupies an intermediate position between zones;

In - % of hepatocytes containing substrate more than 75% of the area of the cytoplasm adjacent to the portal tract,

and when the value of R & d equal to 0.37 and below, determine the response of the stress-workout, and a value of IL more than 0.7 determine the reaction activation.



 

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