Method of treating hyperparathyreosis

FIELD: medicine.

SUBSTANCE: method of treating hyperparathyreosis involves patient's blood sampling prior to a surgery by excision of parathyroid adenoma vessels, removal of the adenoma, blood sampling in 5 and 10 minutes after excision of an adenoma vascular pedicle, parathyroid hormone ELISA test involving centrifugation of the samples, cultivation of the prepared centrate in plates containing ELISA reagents, ELISA interruption before a stabilisation phase by washing of the and adding chromogen then, evaluation of optical density of the solutions containing patient's blood serum, with considering a successful surgery and stating no need of bilateral neck cavity exploration required in observing the decrease of optical density of the solution sampled after adenoma resection in comparison with a pre-operative sample more than twice; the blood samples are centrifuged for 30-40 sec; the blood samples are divided into a number of portions and placed in different plate wells containing the ELISA reagents, incubation interruption and optical density analysis of the solution are performed simultaneously in three different samples making a series, and incubation time of the first series makes 5 minutes, and duration of incubation of the following series differs by 1 minute, and a success criterion is a relative decrease of optical density of the pre- and post-operative solutions. The analysis result is considered to be negative in the absence of the decrease of optical density of the pre- and post-operative solutions in the samples 10 minutes after of blood sample cultivation.

EFFECT: decreasing intraoperative injuries and improved cosmetic effect herewith maintaining a high level of operational efficiency and considerably reduced operation length.

2 cl, 1 ex

 

The invention relates to medicine, more specifically to methods of treatment of diseases of the parathyroid glands, namely hyperparathyroidism.

Currently, the most widespread method for the treatment of hyperparathyroidism associated with the formation of the patient adenomas of the parathyroid glands, is a bilateral surgical revision of the cavity of the neck with the visual determination of the state of the parathyroid glands, identifying adenomatosna modified parathyroid gland and removing it. This method is highly efficient and results in most (to 92.8%) cases to normalize the level of parathormone after the operation and restoration of normal parameters of calcium and phosphorus metabolism [Russel C.F., Edis A.J. Br. J. Surg., 1982. - V.69. - P.244-247].

However, bilateral revision of the cavity of the neck is quite traumatic intervention, carried out through the skin incision length of 7-8 cm, and is accompanied by tissue injury of the neck on both sides with subsequent scar formation process in the area of operation. In addition, when it is used, there is increased likelihood of postoperative hypoparathyroidism [Tibblin S., Bondeson A.G., Ljungberg O. Ann. Surg. 1982. - V.195. - P.245-252].

There is a method of treating hyperparathyroidism, which consists in conducting unilateral revision of the cavity of the neck with the visual determination of the state of colomitos is the breaking glands only on one side (right or left) of the [S.I. Roth, Wang C.A, J.T. Potts Hum. Pathol. 1975. - V.6. - P.645-648]. Definition of the side of the body which carried out audit of the parathyroid glands, is made on the basis of the preoperative examination, including ultrasound and subtraction scintigraphy with technetium. Such surgery can achieve cure of hyperparathyroidism in 88.8% of cases, while reducing trauma intervention compared with bilateral revision of the cavity of the neck [Bergenfelz A. et al. Ann. Surg. 2002. - V.236. - R-551].

However, the cosmetic effect of unilateral revision of the cavity of the neck is similar to the effect of bilateral revision. In addition, the probability of persistence of hyperparathyroidism after unilateral revision exceeds the risk after bilateral revision, because visually inspected not all of the parathyroid glands, resulting in a still presence in the body of the patient the second, not identified in the diagnostic phase, the adenoma. The frequency of occurrence of double adenomas of the parathyroid glands is up to 15%, i.e. is sufficiently high [Siperstein A. et al. Surgery. 2004. - V.136. - R-880].

There is a method of treating hyperparathyroidism, which consists in selective parathyroidectomy, i.e. the removal of adenoma of the parathyroid gland without revision of the remaining parathyroid glands [Udelsmn R. Ann. Surg. 2002. - V.235. - P.665-670]. Localization of the affected parathyroid gland is determined by the results of ultrasound and scintigraphy of the parathyroid glands, and removal of adenomas is performed through an incision of small size (2-5 cm), which improves the cosmetic result of the operation and reduces the severity of postoperative pain. However, there is a probability of persistence of hyperparathyroidism after surgery, associated with the possible presence in the body of the second adenoma of the parathyroid gland.

The closest to the technical nature of the claimed method is selective parathyroidectomy, in which the absence of the second adenoma is controlled by changing the level of parathormone serum after 5 or 10 minutes after removal of the adenoma [Bergson E.J. et al. Arch. Otolaryngol. Head Neck Surg. 2004. - V.130. - N.1. - P.87-91]. Measurement of hormone levels carry out enzyme-linked immunosorbent assay, which includes centrifugation of the blood sample within 5 minutes, the room received serum in wells containing fixed to the solid phase antibodies to parathyroid hormone, and conjugate the antibody-enzyme, incubating for 30 minutes. Then the wells are washed, removing the unbound portion of the conjugate, make a solution of substrate (Chromogen)to measure the optical density about the s and determine comparing with the calibration curve, the number of parathormone in the sample in PG/ml or pmol/L. the decline of the level of parathormone in the serum of more than 50% of the initial preoperative level is considered to be evidence of the lack of additional adenomas of the parathyroid glands in a patient, which allows the surgeon does not conduct bilateral revision of the cavity of the neck to determine the status of the remaining parathyroid glands.

The disadvantage of this method is the length of the laboratory analysis is not less than 40 minutes, including 5 minutes spent on centrifugation of the blood sample, 30 minute - incubation (30 minutes is the stabilization of the formed complexes of the antibody-antigen-antibody-enzyme), 5 minutes by washing the sample, adding Chromogen solution, reading the results of a study comparing the result with the calibration curve and the results in the selected measurement units. In the result, there is a need for long waiting for the results of laboratory studies during the operation, which greatly extends its length.

Technical problem on which this invention is directed, is the reduction of the operation time of patients with primary hyperparathyroidism by conducting selective parathyroidectomy with intraoperative determine the population dynamics of changes in the level of parathormone blood.

The basis of the decision task was based on the problem of finding such a mode of processing blood, which would allow to obtain data about the absence of additional adenomas of the parathyroid glands in a shorter time. During the experiments it was found that technical result can be achieved by carrying out centrifugation for 30-40 with blood samples taken 1 minute before the cut-off vessels adenoma of the parathyroid gland, and after 5 and 10 minutes after the cut-off vessels adenoma with subsequent simultaneous cultivation of each of the samples on multiple wells and simultaneous interruption of the immune response after every 1 minute of starting with 5 minutes in three different samples, forming a series, at different periods of incubation by washing the wells followed by the addition of the Chromogen and the measurement of the optical density of the obtained solutions with the registration of the reduction optical the density of the solution obtained by the processing of blood samples taken after the cut-off vessels adenomas, more than 2 times compared with the solution obtained by processing samples immediately before the cut-off vessels adenomas, the test is considered positive, and bilateral revision of the cavity of the neck is not performed. As a rule, the examination is carried out up to 10 minutes cultivation inclusive. No set is on result within 10 minutes of cultivation is considered as indicating the presence of the second adenomas.

Laboratory investigation of all three blood samples are produced simultaneously, which enables the analysis of the results of a typical calibration curve of the analyzer as the analyzer calibration in the study of all three blood samples is the same, and additionally to reduce the time of getting results, and enable automated analysis of samples.

The proposed method allows to replace the calculation of the level of parathormone in the blood in absolute values for the comparison between the optical density of the corresponding solutions. As from the start of incubation curves of the binding of immune components (antibodies, hormone molecules, complexes of antibody-enzyme") in solutions containing the blood plasma of the patient before the cut-off vessels adenomas and after 5 and 10 minutes after the cut-off vessels, are expressed offset from each other, the difference in optical density after the addition of the Chromogen is fixed reliably at an early period of incubation. In most cases, the drop in optical density 2 times detected already after 4-5 minutes of incubation, and therefore the test result can be obtained much earlier than would be achieved by a stabilization phase (30 minutes).

The drawing shows the differences in the curves of variation of the optical p is h solutions obtained by treatment of the blood plasma of the patient taken before removal of the adenoma (upper curve) and after 5 minutes after its removal (lower curve). As an example, the selected measurement results of the relative density of the solution, expressed in relative light units (RLU), between the test samples on the incubation period of 4 minutes.

The nature and advantages of the proposed method, and industrial applicability of the invention are illustrated by the following examples.

Example 1. In the clinic UHF STMTS University in the period from September 2010 to February 2010 were in the treatment of 35 patients with a diagnosis of Primary hyperparathyroidism". In all patients the localization of adenomas of the parathyroid gland was determined before surgery by using ultrasound and subtraction scintigraphy with technetium.

Patients was carried out surgery in volume selective parathyroidectomy (in 20 cases in the traditional way, in 15 cases by using videoassistance methods of removal of adenomas). The size of the skin incision ranged from 1.5-2 cm in videoaccelerator.com method of operating up to 3-3,5 cm in the traditional intervention.

In 33 cases, the use of preoperative data of the survey allowed us to localize the adenoma of the parathyroid gland in perational wound. After mobilization adenomas, 1 minute before crossing vessels adenomas, conducted the sampling of venous blood of the patient, after which the leg of adenoma was kopirovali and crossed. After 5 and 10 minutes after clipping and crossing vascular pedicles adenomas conducted sampling of venous blood, after which all three samples were simultaneously placed in a centrifuge and centrifuged for 30 seconds at a speed of 3000 rpm.

After 30 seconds of each tube were selected part of the blood serum, which was transferred into wells for enzyme immunoassay and placed in immunoassay analyzer. Incubation of blood samples with standard reagents for immunoassay (time to reach the phase stabilization of the enzyme reaction for a used set of reagents for 30 minutes) was carried out at room temperature in several wells. The immune reaction was interrupted in three samples simultaneously through 2, 3, 4, 5, 7, 8, 9, 10 minutes by rinsing the appropriate wells followed by the addition of the Chromogen solution. Conducted analysis of the optical density of the obtained solutions, expressed in relative light units (RLU, Relative Light Unit). Based on the results of this analysis were decided on the need for bilateral revision cavity neck is whether the termination of the operation. In the fall of optical density more than 2 times in samples obtained after 5 or 10 minutes after removal of adenomas, compared with the sample obtained prior to removal of the adenoma, the test result is considered positive, in connection with which the operation was stopped and had stitches on the wound. In the absence of the dynamics of the decline in optical density or by increasing its held bilateral revision of the cavity of the neck.

The tube with the rest of the blood and deposited on the bottom of the shaped elements continued centrifuged for 4.5 minutes, then was removed from the tubes serum. This part of the serum was placed in immunoassay analyzer, and incubated according to the standard scheme (incubation 30 minutes, rinse, adding Chromogen, reading results, the calculation of the absolute level of parathormone in pmol/l by comparison with a standard calibration curve).

In 17 cases, patients noted a decline in the optical density of the solutions obtained during the processing of blood samples taken after 5 and 10 minutes after removal of the adenoma in 2 times in comparison with the optical density of the solution, obtained by treatment of the blood serum of the patient taken before the cut-off vessels adenomas, when the time of incubation in 4 minutes, in 3 cases - when the time of incubation in 5 minutes, in 3 cases - when the time of incubation in 7 mi the ut, in 7 cases, with the time of incubation for 8 minutes, in 2 cases - when the time of incubation in 10 minutes. A similar result was seen as showing the absence of a second adenoma of the parathyroid gland in a patient, and therefore surgical intervention was limited to selective parathyroidectomy, and bilateral revision of the cavity of the neck was not conducted. All patients in this group after 2 months after surgery remained normal levels of parathormone and ionized calcium blood, testified to the recovery. The average incubation time of the blood sample to obtain diagnostic response test was 5,14 minutes.

In 2 cases after removal of a lesion identified in the area of the expected location of the adenoma, the decline in the optical density of the solutions obtained by treatment of a patient's blood, taken after 5 and 10 minutes after removal of the adenoma was only 10% of baseline after 10 minutes of incubation, and in 1 case it was noted the increasing level of optical density. In these cases there was bilateral revision of the cavity of the neck. In 2 cases after revision cavity neck was revealed adenoma of the parathyroid gland, located near the site of the initial audit, but not the remote surgeon (the tissue that is deleted during the initial audit, subsequent the m histological examination was recognized as a lymph node), and in 1 case the identified second adenoma of the parathyroid gland, located on the opposite side of the neck. After removal of adenomas by bilateral revision of the decline in optical density corresponded diagnostic: in all patients the optical density of the solutions obtained by treatment of a patient's blood, taken after 5 and 10 minutes after removal of the adenoma, under cultivation for 5 minutes decreased 4 times. All patients who underwent bilateral revision cavity neck, 2 months after surgery remained normal levels of parathormone and ionized calcium blood, testified to the recovery.

The survey research of parathormone blood, expressed in absolute figures, showed similar results. In any case, there were no differences between the dynamics of reduction of the level of optical density and dynamics of the lower level of parathormone blood. However, the assessment of the absolute level of parathormone was possible only after 30-40 minutes after taking the sample.

Thus, the inventive method allows to reduce the analysis time while maintaining a high level of efficiency of operations, improve the accuracy of determining the dynamics of the decrease of optical density using a simultaneous laboratory analysis of all three samples is Ravi, that allows you to conduct research in the most similar conditions and the same calibration status of the instrument.

A modified method of treatment to reduce the morbidity of surgical intervention and improve cosmetic results while significantly reducing its duration compared to the prototype; to reduce the probability of persistence of hyperparathyroidism after surgical treatment.

1. A method of treating hyperparathyroidism comprising sampling the patient's blood before the operation, cut-off vessels adenoma of the parathyroid gland, removal of the adenoma, the sampling of blood after 5 and 10 min after clipping vascular pedicles of the prostate gland, the determination of parathormone enzyme-linked immunosorbent assay, comprising centrifuging samples, culturing the obtained supernatant tablets containing reagents for immunoassay, interrupt the immune response to achieve its stabilization phase by washing the wells followed by the addition of the Chromogen, the determination of the optical density of the solutions containing the serum of the patient, followed by a decision about the success of the operation and without bilateral revision cavity neck while reducing the optical density of the solution taken after resection of the adenoma what about the comparison with sample taken before surgery more than 2 times, characterized in that the blood samples are centrifuged for 30-40 C, blood samples are divided into several pieces and placed in different wells containing reagents for immunoassay, interruption of incubation and analysis of the optical density of the solution is carried out in parallel in three different samples, forming a series, and the incubation time of the first episode is 5 minutes and the duration of incubation following series differ from each other for 1 min, and as a criterion of success of the operation using a relative decline in the optical density of the solutions taken before and after resection.

2. The method according to claim 1, characterized that the result of the analysis is negative in the absence of reducing the optical density of the solutions taken before and after resection in the samples after 10 min culturing blood samples.



 

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2 ex

FIELD: medicine.

SUBSTANCE: method involves applying one or two parallel through draining tubes having lateral perforations. Flow lavage of the retroperitoneal space with antiseptic solutions is carried out via the perforations at room temperature and cooled solutions are administered concurrently with vacuum suction. Omental bursa is concurrently drained using the two parallel through draining tubes. Flow lavage of the omental bursa is carried out using these tubes.

EFFECT: enhanced effectiveness of treatment in healing pyo-inflammation foci.

5 cl, 1 dwg

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