etd.page.titleprefix Neurohistocheminio tyrimo panaudojimas diagnozuojant Hiršprungo ligą
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Date
2017
Authors
Kužmarskytė, Rita Skaistė
Journal Title
Journal ISSN
Volume Title
Publisher
Lietuvos sveikatos mokslų universitetas
Abstract
Šio darbo tikslas yra įtariant vaikams Hiršprungo ligą, panaudojant neurohistocheminį tyrimą, ją diagnozuoti.
Darbo uždaviniai: įvertinti padarytus preparatus mikroskopuojant ir nustatyti ar yra matomi nerviniai mazgai tiriamajame storosios žarnos segmente. Taip pat įvertinti ar yra matomos acetilcholinesterazės pozityvios nervinės skaidulos gleivinėje ir aptikus šias nervines skaidulas suskaičiuoti jų tankumą tiriamajame preparate.
Medžiaga ir metodai: tyrimui rektoskopinės biopsijos metu paimti tiriamosios medžiagos ėminiai iš 7 vaikų, kuriems įtariama Hiršprungo liga. Paimta medžiaga greitai užšaldyta panaudojant skystą azotą. Laboratorijoje kriostato pagalba atlikti 15 µm storio pjūviai. Toliau medžiaga buvo dažoma, panaudojant AchE dažymą pagal A. L. Badawi ir E. A. Schenk metodiką. Preparatai buvo mikroskopuojami, lyginami su kontroline medžiaga ir vertinami esami pokyčiai. Atrinkti 52 preparatai, kuriuose matomi skersiniai storosios žarnos pjūviai. Juose buvo skaičiuojamas aptiktų nervinių skaidulų tankumas.
Rezultatai: preparatuose nustatyta, kad paveiktame storosios žarnos segmente, pogleivyje nėra nervinių mazgų. Taip pat stebimas didelis kiekis AchE teigiamų nervinių skaidulų, kurios siekia gleivinę. Tyrimo metu suskaičiuotas šių nervinių skaidulų tankumas, tam tikrame preparato plote.
Išvados: atlikus tyrimą buvo stebimi ligos diagnozei būdingi histologiniai pokyčiai, todėl galima teigti, jog neurohistocheminis tyrimas yra patikimas ir vertingas diagnozuojant Hiršprungo ligą vaikams.
The aim of this thesis is to diagnose children, who are suspected to have Hirschsprung’s disease, using neurohistochemical test. The objects of the thesis: to evaluate made preparations with microscope and to determine whether or not the ganglia is visible in the segment of the large intestine that is under observation. Also, to evaluate whether or not the positive nerve fibers of acetylcholinesterase are seen in mucosa, and to count the density of these found nerve fibers in the preparation under investigation. Material and methods: the samples for the research were taken by the means of rectal biopsy from 7 children, who are suspected to have Hirschsprung’s disease. The taken material was quickly frozen with liquid nitrogen. With the help of cryostat 15 µm thick cuts were made in the laboratory. Next, the material was painted using AchE colouring, according to A. L. Badawi and E. A. Schenk methods. Preparations were observed with microscope, compared with control material and current changes were evaluated. 52 preparations were selected where cross-sectional cuts of the large intestine are visible. There the density of found nerve fibers was counted. Results: it was determined in the preparations that there are no ganglia in the affected segment of the large intestine, in its submucosa. Also, a large amount of AchE positive nerve fibers are under observation, which reaches mucosa. During the research the density of these nerve fibers was counted in the certain area of the preparation. Conclusions: histological changes specific to diagnosis of the disease were observed after the research, therefore, it is possible to state that the neurohistochemical test is reliable and valuable in diagnosing Hirschsprung’s disease for children.
The aim of this thesis is to diagnose children, who are suspected to have Hirschsprung’s disease, using neurohistochemical test. The objects of the thesis: to evaluate made preparations with microscope and to determine whether or not the ganglia is visible in the segment of the large intestine that is under observation. Also, to evaluate whether or not the positive nerve fibers of acetylcholinesterase are seen in mucosa, and to count the density of these found nerve fibers in the preparation under investigation. Material and methods: the samples for the research were taken by the means of rectal biopsy from 7 children, who are suspected to have Hirschsprung’s disease. The taken material was quickly frozen with liquid nitrogen. With the help of cryostat 15 µm thick cuts were made in the laboratory. Next, the material was painted using AchE colouring, according to A. L. Badawi and E. A. Schenk methods. Preparations were observed with microscope, compared with control material and current changes were evaluated. 52 preparations were selected where cross-sectional cuts of the large intestine are visible. There the density of found nerve fibers was counted. Results: it was determined in the preparations that there are no ganglia in the affected segment of the large intestine, in its submucosa. Also, a large amount of AchE positive nerve fibers are under observation, which reaches mucosa. During the research the density of these nerve fibers was counted in the certain area of the preparation. Conclusions: histological changes specific to diagnosis of the disease were observed after the research, therefore, it is possible to state that the neurohistochemical test is reliable and valuable in diagnosing Hirschsprung’s disease for children.
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Keywords
Hiršprungo liga, Neurohistocheminis tyrimas, Acetilcholinesteraze., Hirschsprung’s disease, Neurohistochemical test, Acetilcholinesterase.