RU2263913C2 - Method for predicting gestosis - Google Patents

Abstract

FIELD: medicine, laboratory diagnostics.

SUBSTANCE: the present innovation deals with predicting gestosis development in the 3d trimester of pregnancy. At the terms of 6-13 wk of gestation one should detect the activity of cathepsin D in peripheral venous blood and at its value being either equal or above 0.021 U/h one should predict gestosis development in the 3d trimester. The innovation provides prediction of gestosis development since the early terms of pregnancy.

EFFECT: higher accuracy of prediction.

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Description

The invention relates to medicine, namely to obstetrics and gynecology, and can be used to predict the development of gestosis in the 3rd trimester of pregnancy in women when examined at 6-13 weeks of gestation.

Gestosis remains one of the most urgent problems of modern obstetrics due to its wide prevalence (up to 42%), the complexity of etiopathogenesis, and the insufficient effectiveness of treatment and prophylactic measures (9). In recent years, there has been an increase in severe forms of gestosis, which are the reason for the increase in perinatal losses and high rates of maternal mortality. The urgency of the problem is also due to the serious consequences of gestosis. Most women who have had this complication develop chronic kidney disease, hypertension, and endocrine disorders (4).

The pathogenetic mechanisms of this pathology are laid already in early pregnancy and only delivery is an effective treatment method (10). Prediction of gestosis allows you to determine the most rational tactics for managing such pregnant women, use all possible preventive and therapeutic measures. In this regard, the development of methods for predicting and preventing gestosis is a complex clinical problem.

A known method for predicting late toxicosis by determining the level of lipids and phospholipids in the urine at a gestational age of 8-12 and 20-24 weeks (5). At lipid levels 513.1 ± 65.8 and 706.4 ± 84.7 units. optical density and phospholipids 5.74 ± 1.7 and 16.5 ± 6.1 mg / l gestosis is predicted. The accuracy of the method is 64.5%.

The method has disadvantages:

- the need for a double examination of a pregnant woman in 8-12 and 20-24 weeks of pregnancy;

- an unsuccessful choice of the research substrate, since the appearance of lipids and phospholipids in the urine is characteristic of chronic diseases of the urinary system (nephritis and pyelonephritis) and other somatic pathology, which can be combined with gestosis, and can be an independent disease;

- low accuracy;

- at present, in modern obstetrics there is no such thing as “late toxicosis”, since the etiological and pathogenetic mechanisms of gestosis develop from the early stages of gestation (8).

Also known is a method for predicting gestosis by determining a membrane-cell index (2). The erythrocyte resistance to peroxide hemolysis is used as a membrane cell indicator, and with its value less than 150 s the development of gestosis is predicted.

The method has disadvantages:

- conduct a study and gestational age, in which the development of gestosis is predicted;

- the accuracy of the method is not indicated;

- the possibility of changing the membrane cell indicator of red blood cells not only with gestosis, but also in the presence of a woman's hematological diseases (hemolytic anemia, including hereditary and autoimmune genesis) in the absence of gestosis;

- the invention does not indicate the gestational age at which the examination should be carried out.

Closest to the claimed method according to the technical solution is a method for predicting late toxicosis of pregnant women by the determination of superoxide dismutase activity in blood serum. With an activity value of more than 5.1 U / ml, the development of late toxicosis is predicted (1).

The specified method is selected as a prototype.

The disadvantages of the prototype method are:

- at present, in modern obstetrics there is no such thing as “late toxicosis”, since the etiological and pathogenetic mechanisms of the development of gestosis are formed from the early stages of gestation (8);

- the invention does not indicate the gestational age in which the study should be conducted, and the gestational age, in which the development of toxicosis is predicted;

- the method is not specific for predicting gestosis, since the activity of superoxide dismutase in blood serum increases also in other pathological processes associated with increased free radical oxidation, for example, with hypertension, nonspecific aortoarteritis, etc. (7);

- determination of the enzyme superoxide dismutase presents certain difficulties in connection with its short half-life.

These disadvantages can be eliminated in the proposed method.

The claimed technical result is achieved by the fact that at 6-13 weeks of gestation in the peripheral venous blood of pregnant women determine the activity of cathepsin D and with values of this indicator equal to or more than 0.021 Ed.act.f./h (units of enzyme activity per hour), they predict the development of gestosis in the 3rd trimester of pregnancy with an accuracy of 80.64%, sensitivity 84.44% and specificity 70.58%.

The novelty of the proposed method is that for the first time it is proposed to use the determination of the activity of cathepsin D in the peripheral venous blood of pregnant women at a gestational age of 6-13 weeks in order to predict the development of gestosis in the 3rd trimester of pregnancy. Cathepsin D is a lysosomal enzyme of the endopeptidase class (3). The activity of cathepsin D, like other lysosomal enzymes, is a reflection of the processes of free radical oxidation in the body, the pathological activation of which is one of the triggers for the development of gestosis (1, 6). Also, to date, the participation of cathepsin D in the processes of apoptosis and placentation has been proven (11, 12). A previous study of cathepsin D activity in pregnant women with gestosis has not been conducted. It is assumed that a change in cathepsin D activity in early pregnancy reflects the development of pathological processes that further contribute to the development of gestosis.

The method is carried out according to the standard method as follows.

0.25 ml of a pH 3.3 acetate buffer and 0.25 ml of a substrate solution (1% hemoglobin) are added to 0.5 ml of peripheral venous blood. Incubation is carried out at 45 ° C for 60 minutes The reaction is stopped by the addition of 5.0 ml of 3% trichloroacetic acid. After 5 minutes, the mixture was filtered through a paper filter (Whatman No. 1 or No. 3) and the E 280 of the transparent filter was measured on a spectrophotometer. In control embodiments, the blood is mixed with an incubated solution of substrate and buffer immediately before the addition of trichloroacetic acid. Enzyme activity is calculated by the difference E 280 between the experimental and control variants. The value in units of optical density (unit op.pl.) at a wavelength of the spectrophotometer 280 is equal to the unit of enzyme activity per hour (ED.act.f./h) (3).

Significant differences of the claimed method are:

- a new prognostic parameter of cathepsin D activity in the peripheral venous blood of pregnant women is proposed, equal to or more than 0.021 Ed.act.ph./h (enzyme activity units per hour), to predict the development of gestosis in the 3rd trimester, starting from 6 to 13 weeks of pregnancy .

The essence of the proposed method is illustrated by the following examples.

Example 1. Pregnant A., 28 years old. Real pregnancy is the first. In the 1st trimester of pregnancy, a woman was hospitalized for early mild gestosis. When examining the claimed method in a gestational age of 7 weeks: the activity of cathepsin D in the peripheral venous blood is 0.021 Ed.act.ph./h, which is equal to the prognostic level for the development of gestosis, the development of gestosis is predicted. When observing during pregnancy, the blood pressure in a woman remained within 100 / 60-120 / 80 mm Hg, the total weight gain was 15.7 kg, and uneven weight gain was noted from 24 weeks up to 1 kg per week. From extragenital pathology during pregnancy, vegetovascular dystonia of a hypotonic type was detected, a high degree of myopia. From 35 weeks of pregnancy, the woman developed swelling of the lower extremities. At 37 weeks she was hospitalized with a diagnosis of Pregnancy 37 weeks. Mild gestosis. Pelvic presentation of the fetus. In the hospital, blood and urine tests revealed no pathology. The scoring of the severity of gestosis according to G.M.Savelyeva was 5 points, which corresponds to a mild gestosis (10). Pregnancy ended in timely operative delivery (regarding the pelvic presentation of the fetus in a primiparous woman) and the birth of a live full-term baby weighing 3600 g, height 51 cm and an Apgar score of 8/9 points. Conclusion: the forecast for the claimed method was confirmed.

Example 2 Pregnant K., 23 years old. Real pregnancy 3. A history of 2 medical abortions at the request of the woman. When examining the claimed method on an outpatient basis during pregnancy for 11 weeks: the activity of cathepsin D in the peripheral venous blood is 0.049 Ed.php / h, which corresponds to the prognostic level for the development of gestosis, the development of gestosis is predicted. When observing during pregnancy, the total weight gain was 14.7 kg, from 27 weeks of pregnancy pathological weight gain was noted - up to 2 kg per week. From extragenital pathology during pregnancy, vegetovascular dystonia, diffuse enlargement of the thyroid gland of the 1st degree, euthyroidism were revealed. From 30 weeks of pregnancy, a woman has the appearance of edema of the legs, a periodic increase in blood pressure to 130/80 mm Hg. Gestosis therapy is performed on an outpatient basis without a significant effect. From 34 weeks of pregnancy, edema of the anterior abdominal wall joins. At 37 weeks she was hospitalized with a diagnosis of Pregnancy 37 weeks. Gestosis of moderate severity. Fetoplacental insufficiency. Fetal development retardation syndrome 1 degree. Chronic intrauterine hypoxia of the fetus. In the hospital, there is an increase in blood pressure to 145/100 mm Hg. Urinalysis revealed proteinuria up to 0.132 g / l. According to an ultrasound study revealed a syndrome of delayed fetal development of 1 degree. The scoring of the severity of preeclampsia by G. M. Savelyeva was 10 points, which corresponds to preeclampsia of moderate severity (10). Pregnancy ended in timely delivery and the birth of a live full-term baby weighing 2900 g, height 52 cm and an Apgar score of 7/8 points. Conclusion: the forecast for the claimed method was confirmed.

Example 3. Pregnant M., 24 years old. Real pregnancy 1. In the period of 9-10 weeks she was hospitalized for early gestosis of mild severity. When examining the claimed method in a gestational age of 10 weeks: the activity of cathepsin D in the peripheral venous blood is 0.007 Ed.act.f./h, which is less than the prognostic level for the development of gestosis, the development of gestosis is not predicted. During the observation of extragenital pathology revealed vegetative-vascular dystonia by hypotonic type. The development of gestosis in a woman during pregnancy, childbirth and the postpartum period was not detected.

Pregnancy ended in timely operative delivery (regarding the pelvic presentation of the fetus in a primiparous woman) and the birth of a live full-term newborn weighing 3400 g, 54 cm in height, and an Apgar score of 8/9 points. Conclusion: the forecast for the claimed method was confirmed.

In this way, 62 women were examined at 6–13 weeks of gestation.

The research results are summarized in table 1.

Table 1 Indicator The number of examined Total women 62 true positive result 38 false positive result 5 true negative result 12 false negative 7 Total:
the accuracy of the claimed method is 80.64%
the sensitivity of the claimed method is 84.44%
the specificity of the claimed method is 70.58%

The advantages of the proposed method

1. A study of the peripheral venous blood of a woman is carried out in the period of 6-13 weeks of pregnancy, which allows to predict the development of gestosis in the 3rd trimester before its clinical manifestations and to carry out a set of preventive measures in a timely manner.

2. Determined the activity of cathepsin D in the peripheral venous blood of pregnant women in the 1st trimester, equal to or more than 0.021 Ed.act.f./h (units of enzyme activity per hour), in which the development of gestosis in the 3rd trimester is predicted.

3. High accuracy of 80.64%, sensitivity of 84.44% and specificity of 70.58% of the method.

Thus, the claimed method is not technically complicated, does not require expensive equipment and reagents, can be performed by a laboratory assistant without a higher medical education, not only in stationary but also in outpatient settings and allows high accuracy to predict from early pregnancy (from 6 to 13 weeks) ) the development of gestosis in women. Conducting research on this method will identify women with a high risk of developing gestosis and timely conduct the necessary complex of treatment and preventive measures that will help reduce the frequency of this pregnancy complication, especially its severe forms, the risk of developing pathology of the fetus and newborn and reduce the economic costs of treatment .

LIST OF REFERENCES

1. Bonarenko S.A., Burlev V.A., Grebennikov E.P. A method for predicting late toxicosis of pregnant women. Copyright certificate No. 1612265.

2. Valeeva P.M., Gaysin I.R., Leshchinsky I.A., Kochergin E.V. A method for predicting gestosis. Copyright certificate No. 2191384.

3. Dingle J. Lysosomes. Research Methods. Mir Publishing House, Moscow 1980; p.342.

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Claims (1)

A method for predicting gestosis by examining the peripheral venous blood of women, characterized in that in the period of 6-13 weeks of gestation, the activity of cathepsin D is determined and, with its value equal to or more than 0.021 Ed.act.f / h, the development of gestosis in 3 trimesters is predicted.