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GASTROENTEROLOGY
INVENTION
Patent of the Russian Federation RU2170935

METHOD OF DIFFERENTIAL DIAGNOSTICS OF DESTRUCTIVE CHANGES IN ACUTE CHOLECYSTITE

METHOD OF DIFFERENTIAL DIAGNOSTICS OF DESTRUCTIVE CHANGES
IN ACUTE CHOLECYSTITE

The name of the inventor: Gribanov GA; Golubev AA; Skripnichenko OV; Rasul Sh.H.
The name of the patent holder: Tver State Medical Academy
Address for correspondence: 170642, Tver, ul. Soviet, 4, Tver Medakademiya
Date of commencement of the patent: 2000.04.14

The invention relates to medicine, in particular to internal diseases and surgery. The method provides high accuracy of differential diagnosis. Autolytic changes in blood serum are determined, and after the incubation immediately and after 4, 12 and 24 hours, the amount of glucose, creatinine and lactate dehydrogenase activity is determined and when the amount of glucose and creatinine decreases to 35 and 30%, respectively, and the lactate dehydrogenase activity increases by 1.5 Times diagnose the destructive form of cholecystitis.

DESCRIPTION OF THE INVENTION

The invention relates to medicine, namely, to surgery and clinical laboratory diagnostics.

The problem of choosing the tactics of treatment of patients with acute calculous cholecystitis remains still topical. The surgeon often faces the choice: - to operate the patient at the height of the attack, when the use of modern minimally invasive methods of treating cholecystitis - laparoscopic cholecystectomy, cholecystectomy from mini-access involves significant technical difficulties, or is impossible at all - or to perform conservative treatment, Allows you to postpone the operation and perform it under more favorable conditions.

In this regard, it seems relevant to develop new ways to assess the severity of destructive inflammatory changes in the gallbladder.

In surgical practice, the diagnosis of acute cholecystitis is usually made according to anamnesis and physical examination. From laboratory methods of diagnosis, a clinical blood test is used, where the indicator of the disease is the number of leukocytes. In typical cases, the number of leukocytes is 10 - 15 × 10 9 / l with a shift of the blood formula to the left. And bilirubin (50 g / l) is slightly increased in 45% of patients and transaminase (5 times) in 25% of patients [1,2,3,4].

In the authors' opinion, the analog of the method we propose is a general clinical blood test. But this method has drawbacks, the number of leukocytes varies under the influence of seasonal, climatic, physiological states of the organism, and in a variety of pathologies, which indicates a low information content of this method and the impossibility of evaluating the degree of inflammatory destruction of the gallbladder with it.

The authors propose an original method of differential diagnosis of the severity of inflammatory, destructive changes in acute cholecystitis - autolytic changes in blood serum.

A method of autolytic changes in the tissues of the medulla of white rats was proposed by GA Kuznetsov. Gribanov [6] and is used by the authors as a prototype.

The metabolism of biochemical compounds is characterized by a combination of processes of their synthesis and disintegration. And one of the most widespread biological phenomena, where decay reactions tend to prevail over synthetic ones, is autolysis [5].

Currently, thanks to the use of new, modern methods for analyzing various biochemical components, and on the basis of numerous studies of autolytic processes in various organs and tissues and their ultrastructures [6,7], new views and approaches to the autolysis process have been developed, a new definition of the process Autolysis. The autolysis process is defined as "the property of biological objects developed in the course of evolution to decompose enzymatically and non-enzymatically by proprietary structures of different levels with the predominant participation of hydrolase as well as transferase and other biodegradation and biotransformation reactions of their molecular components" [6].

To study the autolysis process, serum from healthy and sick people with a clinically established diagnosis of acute cholecystitis was used, with a detailed picture of the disease confirmed by special studies. As a control, studies were conducted in practically healthy people. A total of 56 men and women between the ages of 40 and 60 were examined.

Autolytic processes with acute cholecystitis with different degree of inflammatory destruction of the gallbladder were considered in the work with two groups with insignificant and strong destructive changes.

Blood sampling was performed before the operation, from the ulnar vein, on an empty stomach. Serum was separated by centrifugation at 3000 rpm for 15 minutes. Samples of blood serum (in a volume of 0.5-1.0 ml) were incubated in sterile tubes in a thermostat at 37 ° C.

Biochemical parameters were evaluated in the following terms: immediately after the sample was obtained - 0 hours, 4, 12, 24 hours after incubation in a thermostat. The choice of these terms of study is due to differences in the nature and intensity of the autolytic process in dynamics.

To assess the changes in the autolytic process, the following biochemical parameters of blood serum were studied: glucose, total protein and fractions, seromucoids, thymol test, urea, creatinine, uric acid, cholesterol, triglycerides, b- lipoproteins and a number of enzymes: aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, Lactate dehydrogenase and its thermostable subunit, a -amylase U- glutamate-transferase and cholinesterase.

Biochemical studies were conducted on unified clinical and biochemical laboratory methods [8] using a semi-automatic analyzer FP-900.

Determination of glucose.

The determination was made by the Biocon kits. Principle of the method:



Preparation of reagents: a mixture of lyophilized reagents is dissolved in 50 ml of distilled water before use and left to stabilize for 30 minutes. The standard solution is ready for use, the concentration is 5.55 mmol / l.

Analysis: 0.9 μl of the working reagent is added with 10 μl of serum or standard solution, mixed and incubated for 25 minutes at 37 ° C., photometry is carried out at 500 nm, the blank sample is measured by reagent.

Definition of creatinine.

The determination was carried out by the "Biocon"

Principle of the method (modified method of Jaffe): creatinine together with picric acid in an alkaline medium forms a colored complex. The rate of coloration is directly proportional to the creatinine content in the sample.

Preparation of reagents: a saturated picric acid solution containing 0.01% detergent is mixed with a 2.5 M NaOH solution. The standard creatinine solution is 200 μmol / L in 0.1 mol / l HCl.

Analysis: 0.5 ml of the working reagent is heated at 37 ° C for 5 minutes, then mixed with a test sample or standard solution in an amount of 25 μl and measured.

Definition of LDH.

The definition is carried out by sets of "LDG-P" of the company "Biocon". Principle of the method:



The reagent is dissolved in bidistilled water and aged for 30 minutes to stabilize at room temperature. The reaction is carried out at 37 ° C, a kinetic reaction with a retardation time of 30 seconds, a reaction time of 90 seconds, at 340 nm 500 μl of the reagent is heated for 5 minutes, 20 μl of the sample is added, mixed and the reaction is started.

As a result of the research, the most significant criteria for assessing the nature of autolytic changes for acute cholecystitis were found, including glucose, uric acid, urea, creatinine, alkaline phosphatase, lactate dehydrogenase. All other investigated biochemical parameters did not give significant deviations in the dynamics of autolysis and were not considered in the future.

In the process of autolysis, a stable pattern of changes in the above biochemical parameters was revealed in comparison with the control group of healthy individuals, depending on the degree of destructive changes in the gallbladder. However, only changes in glucose (carbohydrate metabolism), creatinine (nitrogen exchange), and changes in lactate dehydrogenase activity are presented for consideration (Figures 1, 2, 3).

During the autolysis of blood serum in the control group (healthy individuals), the deviations of the investigated parameters were insignificant, while in patients the deviations from the initial data were more pronounced. For example, during the study of autolytic changes in glucose in patients with nondestructured gall bladder, it was found that the changes were similar to changes in the control group. And in patients with severe inflammatory destruction, the serum glucose level decreased by the end of the study period by 35% (Fig. 1). When studying changes in creatinine content, it was found that in the acute catarrhal form of cholecystitis the dynamics of creatinine content is similar to the dynamics of the control group (gradual increase by the end of the incubation period in the control group by 10% and 18% in patients with catarrhal cholecystitis), and in destructive forms of cholecystitis And the creatinine content decreased by almost 30% compared to the baseline level (FIG. 2). Investigation of the changes in lactate dehydrogenase activity showed that the enzyme activity in autolysis normally decreases, whereas in acute catarrhal cholecystitis it does not change significantly, with destructive forms of cholecystitis increasing the enzyme activity by 1.5 times (Fig. 3).

Thus, it can be concluded that differential diagnosis can be made using the method of assessing autolytic changes in the blood serum of a patient with the degree of inflammatory, destructive change in the gallbladder.

INFORMATION SOURCES

1. Arseniy AK Diagnosis of acute inflammatory diseases of the abdominal cavity. Kishinev. 1982. Стр. 76, 84, 95, 175.

2. Jurg Hegglin. Surgical examination. M., 1980, Стр. 195.

3. Lidsky A.G. Symptomatic diagnosis of surgical diseases. 1973. Стр. 57.

4. Astapenko V.G. A guide to the diagnosis and differential diagnosis of surgical diseases. Minsk, 1988. 96.

5. Lushnikov E.F. Shapiro N.A. Autolysis. - M .: 1974. Стр. 65.

6. Gribanov G.A. Research of biochemical transformations of endogenous lipids of biological structures at autolysis: Dis .... kand. Doctor of Biological Sciences / KSU. - Kalinin, 1986. Стр. 461.

7. Skripnichenko O.V. Gribanov G.A. On the use of autologous serum of patients with ischemic heart disease, myocardial infarction and atherosclerosis with the purpose of biochemical differential diagnosis of TSU. Scientific notes Volume IV. 1999. Стр. 37-39.

8. Menshikov V.V. Laboratory methods of research in the clinic. M .: Medicine, 1987. Стр. 86, 98, 112.

CLAIM

The method of differential diagnosis of destructive changes in acute cholecystitis, including the determination of autolytic changes in blood serum, characterized by the fact that after incubation immediately and after 4, 12 and 24 hours, the amount of glucose, creatinine and lactate dehydrogenase activity is determined, and when the amount of glucose and creatinine is reduced to 35 and 30%, respectively, and an increase in the activity of lactate dehydrogenase in 1,5 times diagnose the destructive form of cholecystitis.

print version
Date of publication 29.03.2007gg