Method of morphometric estimation of prognosis of course of aplastic anaemia after splenectomy

FIELD: medicine.

SUBSTANCE: after the ablation of the spleen, its weight is determined, the average area of a marginal zone of the spleen in histological cuts with coloration with hematoxylin and eosin is measured morphometrically. The obtained values are used to calculate the weight of the marginal zone and, if its values are ≤1.9 g, a favourable prognosis for the course of aplastic anaemia is made, if its value is >1.9 g, an unfavourable course of the disease is predicted.

EFFECT: prognosis of the aplastic anaemia course after splenectomy independent on the severity of the disease with the possibility to approach therapeutic treatment in a differential way.

2 ex


The invention relates to medicine, namely to Hematology, and can be used for morphometric evaluation of the prognosis of aplastic anemia (AA) after splenectomy by determining the mass of the spleen and its marginal zone in the histological sections. There are only few works devoted to the description of the structure of the spleen and changes in its architectonics with AA. In the literature there are no data regarding the influence of the mass of the marginal zone of the spleen in the course of the pathological process in AA. Used high-tech methods of treatment impose more stringent requirements on the search for new criteria for predicting the course of this disease of the blood system, characterized by the substitution of cellular bone marrow fat and, as a consequence, the pancytopenia in peripheral blood.

The aim of the invention is to provide a method of morphometric evaluation of the prognosis of AA after splenectomy by mass of marginal zone of the spleen.

Closest technical solution to the proposed invention is a method of assessing the functional state of components of the white pulp of the spleen with the use of antibodies expressed only on CD20+ and CD3+ lymphocytes, which allows to evaluate the functional status of the white pulp of the spleen in portal hypertension (LiZ., Zhang S., G. Lv et al. / Changes in count and function of splenic lymphocytes from patients with portal hypertension // World J. Gastroenterol. - 2008. - Vol.14 (15). - P. 2377-2382). This method does not allow to accurately assess quantitatively the state of the white pulp of the spleen and its components, time-consuming, costly and requires large amounts of expensive consumables.

Way morphometric evaluation of the prognosis of AA after splenectomy by weight of the spleen and its marginal zone does not exist. In the process of conducting patent information search did not reveal sources to be novelty of the invention.

The invention developed in the laboratory of pathomorphology blood INSTITUTE KNIG and PC FMBA of Russia in accordance with the research plan.

The method is as follows.

The studies were performed on spleen samples from 31 patients with AA aged 18 to 65 years (12 - with a mild form of the disease and 19 with severe). According to the morphometric analysis of the favorable course of the disease found in 11 patients (8 with mild; 3 - severe). In 20 patients identified poor prognosis (16 - in severe and 4 - light). The comparison group consisted of samples of spleens taken from 20 individuals did not have a history of blood system diseases.

After splenectomy determine the mass removed spleen, perform histological wiring samples �sleepering material by the conventional method. Histological sections with a thickness of 3-5 μm, prepared from paraffin blocks spleen, performed morphometric analysis on a light microscope with an integrated photo and video camera from Leica with using software image analysis ImageScope Color, version M with eyepiece ×10, when the lens ×20. In the study of white pulp of the spleen, using the standard method of staining with hematoxylin and eosin, marginal zone in preparations are distinguished by colours. To determine the average size of marginal zone using the morphometric analysis carried out measurements in 50 fields of view for each sample, starting the calculations from the capsule deep into the lymphoid organ. Based on the results obtained, calculate the mass of the marginal zone of the spleen by the formula: M=m×D%/100%,

where M is the mass of marginal zone (g),

m is the total mass of the spleen (g)

D - average size of marginal zone, %.

During the processing of the data was calculated the average mass of marginal zone of the spleen in patients with AA. In the process of conducting research in patients with AA in morphometric histological sections of spleen stained with hematoxylin and eosin to measure the average size of marginal zone and calculate its mass by the formula. The favorable prognosis of AA ascertain when the mass values of marginal zone ≤1.9 g and with values > 1.9 g install unfavorable course of the disease.

The research results presented in the examples, confirming the possibility of the inventive method to carry out morphometric assessment of the prognosis of AA.

Example 1

Patient T., age 21, was admitted to the Hematology unit DIRECTOR KNIG and PC FMBA Russia 19.07.1999 G. blood test on admission: hemoglobin 79 g/l, granulocytes of 0.3×109/l, platelets 15,0×109/l. The diagnosis of aplastic anemia, a severe form. The patient was given treatment by hours worked in the clinic method (patent for invention №2412708 from 27.11.2008. Surgery performed (03.08.99 G.): midline laparotomy, splenectomy - without complications. Weight of the spleen of a patient (m) was 100 g. Conducted a morphometric study of histological samples of the spleen, stained with hematoxylin and eosin. The average size of marginal zone (D) was 2.0%. The mass of marginal zone of spleen (M) - 2.0 g Set value obtained in the following calculation: M=100,0 g×2,0%/100%.

Conclusion: using the data obtained established that patients with an unfavorable prognosis of AA.

Example 2

Patient E., age 39, was admitted to the Hematology unit DIRECTOR KNIG and PC FMBA Russia 06.04.1995 G. the Diagnosis of aplastic anemia, a severe form. Blood test on admission: GE�oglobin 36 g/l, granulocytes of 0.37×109/l, platelets 15,0×109/l. Surgery performed (11.05.95 G.): midline laparotomy, splenectomy without complications. Weight of the spleen was $ 170,0) underwent morphometric evaluation of the histological samples of the spleen, stained with hematoxylin and eosin. The average size of marginal zone (D) was 0.6%. The calculated mass of the marginal zone of the spleen (M) - 1,02 g by the previously mentioned formula. The determined value is obtained in the following calculation: M=170,0 g×0,6%/100%.

Conclusion: using histological data obtained established that patients with a favorable prognosis of AA.

The claimed invention has novelty, gives a positive result consists in the possibility of mass of the spleen, morphometric analysis of the area of the marginal zone and its weight to predict the nature of the disease in patients with AA after splenectomy. The proposed method is simple to implement and can be used in practical Haematology, morphological and histological laboratories as an additional criterion for the prediction of the AA, regardless of the severity of the disease, and allows a differentiated approach when prescribing therapy.

Method morphometric evaluation of the prognosis of aplastic anemia after splenitis�khimii, characterized in that after removal of the spleen determine its mass, morphometric measure the average size of marginal zone of spleen in histological sections when stained with hematoxylin and eosin, given these values, calculate the mass of the marginal zone and its values ≤1.9 g establish a favorable prognosis of aplastic anemia, and if >1.9 g state the unfavorable course of the disease.


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