RU2433410C2 - Method of predicting development of pathological process direction in patients with brain tumours - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: on the first day after operation in liquor activity of proteinase of broad substrate specificity - plasmin is determined. If index increases to 0.200 CE/ml development of tumour recurrence within 4-5 months after operation is predicted and if plasmin activity is over 0.200 CE/ml is recurrence-free course of postoperative period during two years is predicted.

EFFECT: method makes it possible to determine at early stages after operation terms of continuous development of malignant brain tumour.

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Description

The invention relates to medicine, namely to biological research in oncology, and can be used to determine the development of a malignant process after surgical treatment of a brain tumor.

A known method for predicting relapse-free survival by studying matrix metalloproteinases.

(MMP-9) (Serum matrikx metalloproteinase-9 in head and neck squamous cell carcinoma is a prognostic marker. Ruokolainen Henni, Pääkkö Paavo, Turpreenniemi-Hujanen Taina. Int. J. Cancer. 2005. 116, No. 3, p. 422 -427).

The authors determined MMP-9 in the blood serum of patients with squamous cell carcinoma of the head and neck. A high level of serum MMP-9 was associated with low disease-free survival (up to 40% of patients) and, conversely, a low level of serum MMP-9 was associated with good survival (up to 69% of patients). It is believed that the level of serum MMP-9 before treatment can serve as a prognostic factor.

However, this method was not used for malignant brain tumors and is not specific to them.

A known method for determining the protein S-100B in the blood serum of patients with melanoma (Significace of serum S-100B in melanoma patients before and after sentinel node bipsy. Smit Leonie HM, Nieweg Omgo E., Korse CatharinaM., Bonfrer Joahnnes MG, Kroon Bin BR Surg Oncol. 2005. 90, N2, pp. 66-70).

However, such an analysis was not performed for malignant neoplasms of the brain, which makes it impossible to predict continued tumor growth with intracranial gliomas. According to the authors themselves, the use of the test for the early detection of relapse of melanoma gives dubious results. The method is only suitable for detecting an already developed relapse of melanoma.

Both methods were not used in the examination of patients with brain tumors and the results obtained in the study of blood serum cannot be used to predict long-term effects, in particular continued tumor growth.

A known method for the determination of serum thymidine kinase in patients with glial CNS tumors (Neuron specific enolase [NSE] and thymidine kinase [TK] as markhers in biological fluids of brain tineos patients. Boiardi A., Munari L., Silvani A., Solero CL , Bombardieri E. Ital. Y. / Neurol. Sci. - 1990. - 11, N4. S.359-366), which we chose as a prototype. The essence of the method is that serum thymidine kinase is determined and used as a marker of disease recurrence after surgical treatment of patients with glial tumors. The method is based on the fluorimetric determination of thymine in biological material by oxidation with 12.5% bromine water and subsequent neutralization with an alkaline solution of o-aminobenzaldehyde.

However, according to the authors themselves, the specificity of the method for glial tumors is 36%, which, in our opinion, is not enough to use this indicator as a marker of disease recurrence. In addition, during the determination, several intermediate metabolic products are formed (glucose, acetolactic acid, and a number of others), which have close fluorescence and interfere with the determination, which reduces the accuracy of the analysis.

The aim of the invention is the early identification of the timing of continued growth of malignant brain tumors in the postoperative period.

This goal is achieved by the fact that in the patient’s cerebrospinal fluid, the activity of a broad substrate specificity proteinase, plasmin, is determined 1 day after the operation, with an increase of up to 0.200 KE / ml compared to that in the control cerebrospinal fluid, tumor relapse is predicted within 4-5 months after operations, and with plasmin activity above 0.200 KE / ml, a relapse-free course of the postoperative period is predicted for two years.

The invention "A method for predicting the direction of the pathological process in patients with brain tumors after surgery" is new, since it is not known from the level of medical advances in predicting the possibility of continued tumor growth.

The novelty of the invention lies in the fact that plasmin activity is determined in the cerebrospinal fluid of a patient taken on the 1st day after the operation and, when it is increased to 0.200 KE / ml, the development of continued tumor growth is predicted for 4-5 months after surgery. Plasmin activity above 0.200 KE / ml allows predicting a relapse-free course of the postoperative period within 2 years. This method of forecasting is new and makes it possible to take adequate measures in a timely manner to develop pathogenetic therapy in order to prevent the development of continued growth and relapse of a brain tumor.

In open sources of information in Russia, the CIS countries and abroad, no indications of a similar method for predicting the development of relapse of a brain tumor have been found.

The invention is industrially applicable, as it can be reproduced and repeated many times in any clinical laboratory of medical institutions of the healthcare system.

The method is as follows. On the 1st day after removal of the CNS tumor, the patient produced cerebrospinal fluid by lumbar puncture. Selected 1 ml of cerebrospinal fluid and used to determine the activity of plasmin. The determination was carried out by a unified caseinolytic method based on the ability of plasmin to cleave protein substrates in a slightly alkaline medium at 37 ° C. The cerebrospinal fluid was incubated with 2% casein at 37 ° C. The reaction was stopped with 10% trichloroacetic acid (introduced into the control before incubation), the samples were cooled, centrifuged, and the amount of protein products of casein degradation was determined in the supernatant at a spectrophotometer wavelength of 280 nm against the control for reagents. For the calculation, a tyrosine calibration curve was used. The results were expressed in casein units per 1 ml of cerebrospinal fluid (Paschina T.S. et al., J. Vopr. Medical Chemistry, 1976, issue 5.). To implement the method requires 3 hours and the usual laboratory equipment.

As a control, cerebrospinal fluid was used in patients with symptomatic epilepsy, treated with arachnoiditis, benign brain tumors 3 months after surgery, physically healthy with mental retardation.

An example of the application of the method can serve as extracts from medical records.

1. Patient: N., born in 1963. Medical history No. 21492 / c.

He was in the hospital on 10/22/07. until 11/12/07.

Diagnosis (main, concomitant, complications): glioblastoma of the frontal lobes of the brain with growth in the corpus callosum, condition after osteoplastic craniotomy with subtotal removal of the tumor. Clinical group: II, b / s.

Г / а dated October 31, 2007 No. 975763-772 - glioblastoma with isomorphic cell regions, sometimes polymorphic cell with extensive foci of necrosis and hemorrhage.

On admission (general condition, complaints, status): general condition: severe; complaints: headaches; neurologically: cerebral symptoms, frontal psycho-pathological syndrome.

Operation 10.24.2007 - osteoplastic craniotomy in the frontal area on the right with subtotal removal of the tumor. At 1 day after surgery, the patient received lumbar puncture in order to reduce intracranial pressure. Liquor is colorless, transparent; 1 ml on the same day was used to determine plasmin. Result: 0.093 KE / ml, a conclusion is drawn on the development of continued tumor growth in the next 4-5 months.

At discharge: the general condition of the patient is closer to satisfactory.

The patient died 4 months after surgery as a result of continued tumor growth.

2. Patient: K., born in 1959. Case history No. 303 / h.

He was in the hospital from 02/04/08 to 02/29/08.

Diagnosis (primary, concomitant, complications):

Dysembryoplastic neuroepithelial tumor of the right frontal lobe of the brain, condition after osteoplastic craniotomy in the frontal region on the right with the removal of the tumor. Clinical group - II, b / s.

G / a from 12.02.2008, No. 7822-7827 - dysembryoplastic neuroepithelial tumor.

On admission (general condition, complaints, status): general condition: satisfactory; complaints: headaches; neurologically: cerebral symptoms, episindrome.

Operation 02/12/2008 - osteoplastic craniotomy in the frontal region on the right with the removal of the tumor. At 1 day after surgery, the patient received a lumbar puncture in order to reduce intracranial pressure and in connection with complaints of a headache. Liquor is colorless, transparent; 1 ml of cerebrospinal fluid was taken to determine plasmin. Result: 0.279 KE / ml, a conclusion is drawn about the relapse-free course of the postoperative period.

At discharge: the general condition of the patient is satisfactory.

It is observed within 1 year, there is no evidence in favor of relapse.

We examined 7 patients with brain tumors who were treated in the CNS tumor department of the Federal State Research Institute of Surgery and underwent surgery to remove a malignant brain tumor.

It was found that in patients with central nervous system tumors after surgical treatment in cerebrospinal fluid, taken 1 day after surgery, the value of plasmin activity index was 1 day from 0 to 0.093 KE / ml. In only 1 patient with a result of “0” on the second day, the number increased to 0.186 KE / ml against the value determined in the control cerebrospinal fluid - 0, which was accompanied by the development of continued tumor growth in these patients over the next 5 months (3-5 months after operations). In patients with a relapse-free course of the postoperative period, plasmin activity was higher and reached 0.465 KE / ml, on average 0.264 KE / ml.

The technical and economic effectiveness of the method lies in the possibility of early prediction of the development of a relapse of the malignant process in the brain, which contributes to the timely implementation of adequate preventive and therapeutic measures.

The specificity of the method for predicting relapse is 100%, for predicting a relapse-free course of the postoperative period is 80%.

Claims (1)

A method for predicting the development of the direction of the pathological process in patients with malignant brain tumors, including surgery, spectrophotometric studies, characterized in that on the 1st day after surgery, the activity of proteinase of broad substrate specificity, plasmin, is determined, with an increase of up to 0.200 KE / ml they predict the development of tumor relapse within 4-5 months after surgery, and with plasmin activity above 0.200 KE / ml, a relapse-free course of postoperative IRS for two years.