How to collect urine analysis for atypical cells. Urinalysis for cytology

The purpose of the study is to diagnose potentially extremely dangerous, life-threatening tumors of the urinary tract of high malignancy. (HGUC). Cytological examination was introduced into world medical practice in 1945 by J. Papanicolaou.

The urinary tract is anatomical structures that perform the function of urination. It consists of the renal pelvis, ureters, bladder and urethra. The transitional epithelium or urothelium covering the urinary tract under certain circumstances becomes the source of neoplasms, which are very diverse, both in morphological structure and in degree of malignancy. Primary malignant tumors of the renal pelvis and ureter make up only 5-6% of all urothelial tumors, so the main share of life-threatening pathology occurs in bladder tumors (Ministry of Health of the Republic of Belarus State Institution RNPC OMR named after N.N. Aleksandrov. “Algorithms for diagnosis and treatment malignant neoplasms", 2012).

Every year in the Republic of Belarus, 1000-1200 new cases of bladder cancer are detected at various stages of development. It is predominantly the male population that suffers. In European countries, in the structure of general cancer incidence, bladder cancer ranks 4th in men and 14th in women. In the overwhelming majority of cases, this is high-grade urothelial carcinoma (HGUC), which manifests itself clinically as hematuria, i.e. the appearance of blood in the urine. Over the past 20 years, the incidence rate in our country has almost doubled. The relatively high mortality-to-morbidity ratio of 0.33 means that every third patient dies, which undoubtedly indicates the lack of effective non-invasive methods for early diagnosis.

In recent years, scientists have been considering the possibility viral nature bladder cancer. Human polyomavirus 1, better known as polyoma-BK virus, has been attracting the attention of researchers in this field for many years as a potentially dangerous factor in the development of neoplastic processes in the urinary system. This virus is widespread throughout the human population. We can say that up to 90% of the adult population of our planet is infected with it. The first strain of human polyomavirus was isolated from urine back in 1971. In the name of this strain, the capital Latin letters BK correspond to the initials of the patient in whom it was discovered. And over the past two decades, our understanding of the role of this virus has seriously evolved. It is undeniable that with a decrease in immunological protection, namely during cytostatic therapy in patients with allograft kidney transplantation, polyoma BK virus is the main cause of BKV allograft nephropathy, urethral stenosis and hemorrhagic cystitis (Drachenberg CB, Hirsch HH, Ramos E , Papadimitriou JC (2005).

The virus enters the human body through the upper respiratory tract and persists for a long time asymptomatically, mainly in the urothelium, as well as in the bone marrow, causing latent infection. A healthy immune system controls the asymptomatic presence of the virus in the human body. However, when immune surveillance is impaired, which occurs for one reason or another, due to suppression of immune function, the BKV virus is activated. And the morphological reflection of the transition from a latent state to an active one is the appearance of specific cells. In the urothelial epithelium, viral particles form intranuclear inclusions, as a result of which the cell acquires very characteristic morphological features. The first description of such cells with inclusions of viral particles was given by Dr. Koss LG 40 years ago, having discovered them in the urine of a patient after a kidney transplant. He called them Decoy cells. They can be of 4 types, and until recently, the prevailing idea among cytopathologists was that these infected cells cannot be classified as malignant. But their appearance in the urine of a patient with allograft transplantation is the earliest sign of the threat of BKV allograft nephropathy. These figures show 4 morphological types of Decoy cells.

In other words, Decoy cells are the earliest precursors of transplant rejection. But only with the introduction into practice of liquid technologies for preparing a drug from freshly released urine or from a bladder washout, with Papanicolaou staining, did it become possible to detect these cells and distinguish them from malignant ones with high sensitivity and specificity. Currently detectionUrinary decoy cell is a screening test for BKV nephropathy. In addition, in patients who have undergone kidney transplantation, along with Decoy cells, urothelial cells are often found with signs of severe cellular atypia, corresponding to the cytomorphological characteristics of high-grade urothelial carcinoma, as well as cells that simultaneously carry features inherent in Decoy -cells, and HGUC cells. These cells not only have mixed features of Decoy cells and malignant cells, but also carry proteins (SV-40 T) of the polyoma BK virus (Galed-Placed I, Valbuena-Ruvira L. Diagn Cytopathol.2011 Dec;39(12 ):933-7). Many of the clinical cases described in the literature indicate a possible oncogenic role of polyoma BK virus in the occurrence of urothelial carcinoma (Hassan S, Alirhayim Z, Ahmed S, Amer S. Case Rep Nephrol.2013; 2013:858139). BKV proteins have a damaging effect on the synthesis of tumor suppressor proteins (p53), which leads to disruption of DNA repair processes. This pathogenetic mechanism underlies the occurrence of genetically unstable changes leading to the transformation of normal urothelium first into dysplastic and then into high-grade urothelial carcinoma (HGUC). HGUC is characterized by a high rate of relapse and progression to muscle-invasive stages T2, T3, T4 with lymph node metastases. HGUC is diagnosed in 95% of all bladder cancer deaths. So, the advantages of cytological examination of the urothelial tract:

Non-invasiveness of the procedure (analysis from a “jar”);
High specificity for diagnosing HGUC, approaching 100%;
High sensitivity for diagnosing HGUC ~ 80%;
~30-70% of patients with HGUC survive;
~5-15% of HGUC cases progress; US statistics.
535,000 bladder cancer patients survived.

The preferred technology for urine cytology preparation is liquid sedimentation (BD SurePath), which we use in our laboratory. Thin-layer smears allow further diagnosis using the entire arsenal of molecular genetic methods (immunocytochemistry, FISH, etc.).

The cytological diagnosis of HGUC combines all genetically unstable malignant urothelial tumors, which are histologically very diverse. In order to optimize approaches to the interpretation of urinary cytopathology and standardize diagnostic criteria, a new cytological classification system was adopted at the international congress in Paris in 2013. It is very similar to the Bethesda system for cytological analysis of thyroid and cervical canal pathology.

Categories	Paris system for grading urinary tract cytology	Probability of malignancy
I	Unsatisfactory/non-diagnostic material	0-10%
II	Negative for high-grade urothelial carcinoma (NHGUC)	0-10%
III	Atypical urothelial cell (AUC)	8-35%
IV	Suspicion of high-grade urothelial cancer (SHGUC)	50-90%
V	Low-grade urothelial carcinoma (LGUC)	~10%
VI	High-grade urothelial carcinoma (HGUC)	>90%
VII	Other tumors, primary and secondary	>90%

So, cytological examination of urine using liquid smear preparation technologies and using the diagnostic criteria of the 2014 Paris Classification is the main non-invasive method for diagnosing and monitoring high-grade urothelial carcinoma.

Women are often exposed to diseases of the urinary system, which are infectious or inflammatory in nature. This is explained by the anatomical features of the structure of the female pelvis. A visual examination using a cystoscope is not always sufficient to make an accurate diagnosis, so it is advisable to conduct cytological studies.

Bladder cytology is a diagnostic test that allows women to check their bladder for cancer.

This study is prescribed in a situation where there is a suspicion of the presence of a malignant tumor in the organs of the urinary system. Often, patients who have a high probability of detecting cancer cells resort to this method.

Indications for cytological examination

Cytology of the bladder is prescribed for those patients who:

a malignant tumor has already been identified tumor organs of the urinary system, cytological studies in this case make it possible to monitor how the disease progresses;
there is a suspicion of the presence malignant tumors, diagnostics allows to identify cancer;
present blood in the urine, this serves as a signal about the presence of pathology, so the doctor must prescribe a cytological diagnosis;
treatment of the disease is completed, timely diagnosis makes it possible to assess the progress of treatment and avoid relapse diseases.

What is studied through cytology

Cytological examination of the bladder is a study using a microscope of the sediment of a urine sample obtained after centrifugation of the biomaterial. This study is carried out by a histologist in a laboratory setting.

The purpose of cytology is to detect abnormal cells that may be excreted in the urine. Diagnostics makes it possible to accurately identify the presence of a malignant process in the organs of the urinary system.

However, if you receive an unsatisfactory, atypical or suspicious result, you should resort to repeated and additional tests.

Preparation for the procedure

To submit a urine sample for analysis, you should adhere to such general rules as maintaining personal hygiene and using a sterile container designed for collecting biomaterial.

A significant difference in preparation for cytology lies in the time of collection and the amount of material collected.

Urine collection is carried out in the morning, but not immediately after waking up. Biomaterial for analysis is collected approximately 2 hours after the bladder has been emptied of fluid accumulated overnight. If you get up in the morning at 7 o'clock, then it is better to immediately visit the toilet and defecate.

Avoid drinking a lot of fluids as they can dilute the urine. It is recommended to immediately submit the sample to the laboratory in order to obtain a more accurate test result.

If biomaterial needs to be taken from people who lead a sedentary lifestyle or are seriously ill, then a catheter is provided for such situations. A number of hygiene procedures should be carried out: wash the perineum, wipe with a towel, install a catheter and collect a urine sample.

Interpretation of the results of cytological examination

In each laboratory, the result of the study is expressed in terms and units of measurement that are different from those obtained in other laboratories. This depends on the equipment used to conduct the study.

The diagnostic results are provided to the attending physician, who studies them and makes a diagnosis. However, there are generally accepted medical terms that are used in all clinics and laboratories, and have an unambiguous interpretation that is accessible even for independent understanding.

An unsatisfactory result indicates that the study did not identify the number of incorrect cells that would allow an accurate conclusion to be drawn, so the tests should be repeated.

An atypical result indicates that cells of an atypical shape were identified in the submitted samples, but such data do not provide accurate information about whether there are cancers in the organs of the urinary system. Additional tests are required.

A suspicious result serves as the first warning sign. This suggests that suspicious cells were detected, but such neoplasms can also be benign.

A positive result indicates that cancer cells have been detected. Which organ of the urinary system has undergone cancer can be identified using specialist diagnostics. In the future, special treatment is prescribed.

A negative result indicates that the presence of cancer cells in the patient’s body has not been detected.

To diagnose bladder cancer, it is necessary to conduct not only a urine test, but also a study of the entire body as a whole. Then, having all the collected tests in hand, the attending physician diagnoses the patient and selects the appropriate treatment.

The positive aspect of cytological diagnosis is that, unlike other studies, it is carried out quickly (3-5 days), and the results obtained make it possible to begin timely treatment. Often, a doctor may order several such studies to make an accurate diagnosis and establish the fact of oncology.

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Speaking about methods of laboratory diagnosis of early oncological pathology in the bladder, they should immediately be divided into routine methods, such as microscopic and cytological examination of urine sediment, and rather complex biochemical ones, which make it possible to identify certain states of homeostasis, not only characteristic of a specific form of malignant tumor, but and those preceding its occurrence.

We mean a biochemical study of urine, which reveals impaired tryptophan metabolism, manifested by the appearance of its carcinogenic metabolites in the urine.

This can occur both in patients with bladder cancer and in practically healthy people, which indicates a certain predisposition to the occurrence of the disease and is considered one of the endogenous risk factors.

The same can be said about the significance of the increased activity of β-hyaluronidase, a urine enzyme, under the influence of which non-carcinogenic complexes disintegrate in the bladder with the release of the active principle. Thus, a complex consisting of 2-amino-1-naphthol, combined with glucuronic or sulfuric acid in the liver and therefore having lost its carcinogenic properties, under the influence of β-hyaluronidase or urine sulfatase, disintegrates in the bladder with the formation of active 2-amino-1- naphthol, which has a carcinogenic effect on the uroepithelium.

Timely detection of early oncological pathology

Speaking about timely detection early oncological pathology (ERP) in the bladder, one should agree with the opinion of M. Mebel et al. that, despite significant progress in the field of diagnosis, routine methods - examination of erythrocyturia, examination of accessible parts of the urinary tract, rectal palpation - remain the basis for early diagnosis, but are clearly underused. It is enough to point out that in many patients who went to the clinic with non-urological complaints, the presence of red blood cells in the general urine analysis did not serve as a reason for referral to a urologist, neither for general practitioners nor for specialized specialists.

Meanwhile, while conducting a prospective epidemiological study of a contingent of 3,400 practically healthy people in order to identify bladder failure in them, we performed a microscopic examination of urine sediment in 2,143 persons (63.0%), and a cytological examination in 696 (20.5%) and revealed a significant percentage of pathology in people who did not intend to seek medical help, but feel practically healthy.

Hematuria was detected in 18.1% of men in the risk group and 7.9% of controls, in women, respectively, 23.1 and 10.7%. Naturally, all these people need further urological examination, which will allow us to establish the cause of urological pathology, including oncological one. It should be emphasized here that laboratory diagnosis of early oncological pathology in the bladder cannot be an independent method; as a rule, it should be supplemented by a cystoscopic examination with a biopsy.

Indications for cytological examination of urine sediment include all forms of macro- and microhematuria, unclear dysuria, cystalgia, abuse of analgesics, and professional contacts with aromatic amines. Cytological examination is indispensable for bladder tumors of the intraepithelial type - cancer in situ, which cannot be detected endoscopically, as well as for bladder diverticulum cancer, urethral strictures, and low organ capacity.

It should be emphasized once again that this theoretically ideal method cannot be independent either in diagnosing recurrent cancer in the bladder, or, especially, in identifying ROP. According to many authors, this is confirmed by a fairly high percentage of discrepancies between the cystoscopic picture and the results of urine cytology; only in 70.0% of patients with uroepithelial cancer, cytological examination reveals cancer cells.

J. Tostain et al., analyzing 500 cytological studies in 342 patients with uroepithelial tumors, showed that for typical papillomas of the bladder, cytological examination is ineffective; for infiltrative cancers of low malignancy, urine cytology makes it possible to make a diagnosis in 66.0% of cases, for high-grade ones - in 80.0%.

A number of authors (Mansat A. et al., Droese et al.) have established a clear correlation between the degree of cell anaplasia and cytological detection of the tumor. In particular, it was shown that when using the Bergevist classification of I, II, III degrees depending on cell anaplasia, in the first degree the tumor was detected only in 19.0%, in the second - 30.0%, in the third - 70.0%.

There are still no generally accepted cytological methods for studying bladder tumors. Some researchers consider the method of exfoliative cytology valuable in the study of urinary sediment of freshly released urine (Enokhovich V.A., 1966; O.P. Ionova et al., 1972), others prefer aspiration biopsy (Volter D. et al, 1981), others indicate that the best results are obtained by the method of flushing the mucous membrane of the bladder (Lomonosov L.Ya., 1978).

Errors in cytological diagnosis

Errors in cytological diagnosis are determined by several factors: poor material obtained during an inflammatory process or tumor necrosis, a small amount of it, or mild cell atypia. Therefore, naturally, a search is underway for the most rational selection of material for work. Thus, N. Holmguist points out the high degree of detection of bladder cancer (1.2 per 1000) in cytological examination of wet preparations of urine sediment and suggests using this technique for screening.

L.Ya. Lomonosov recommends for mass preventive examinations of high-risk groups the method of active flushing of the bladder in its own modification, when before flushing the bladder is thoroughly washed with furacillin, after which 50-100 ml of an alcohol solution of novocaine is administered at the rate of 15 ml of 96° alcohol per 100 ml of 1% novocaine solution.

After 5-10 minutes, the solution is collected in a clean container. After macro-description of the washout, the material is centrifuged at 3000 rpm for 15 minutes. The supernatant is washed off, and smears are prepared from the centrifuge.

At the same time, a number of authors worked on a rational technique for preparing smears, since the delay in the introduction of cytological examination is largely due to the difficulties of staining cells in urine. Therefore, S. Fiedler et aI. Several stains are carried out - with methylene green, pyronine and chromalone using a method modified by them.

False-positive results are most often associated with incorrect assessment of urothelial cells with dysplastic changes in urolithiasis, a chronic inflammatory process. Thus, of 135 patients with upper urinary tract stones, 7.2% had a cytological picture similar to well-differentiated cancer. In the urine of persons who suffered from urolithiasis for a long time, S. Dimopoulos et al. observed the presence of cells of grade 3 and 4 malignancy according to Pappanicolaou and their disappearance after surgical removal of stones.

O.P. Ionova et al. During inflammatory processes, atypical cells were found in cytograms that differed little from tumor cells. Interpretation of cytograms seems to be quite difficult, which may be why there are still no generally accepted criteria for malignancy of epithelial tumors of the bladder.

The available numerous data on this issue relate mainly to the diagnosis of cancer relapses and quite rare cases of recognition of “cancer in a cell” - cancer in situ. Only a few authors have described cytograms for simple and proliferating papillomas of the bladder - G.A. Arzumanyan, V.A. Enochovich, O.P. Ionova et al., L.Ya. Lomonosov.

A number of others - M. Beyer-Boon et al. indicate that papillomas cannot be diagnosed at all on the basis of cytograms; the latter in typical papillomas do not differ from normograms, and only the detection of papillary tumor fragments allows a diagnosis to be made.

The fact is that cytograms of bladder washouts are normally poor in cellular elements. Most of the cells found are cells of the integumentary zone of the transitional epithelium. They are large in size, polygonal or elongated in shape. The kernels are small, round or oval, located centrally or slightly eccentrically.

The cells of the intermediate zone are cylindrical in shape, arranged either in the form of papillary fern-like structures, or in the form of rosettes, or in the form of grape-like clusters. The nuclei are oval, the chromatin is finely lumpy, evenly distributed over the area of the nucleus. In some studies, attention is drawn to giant multinucleated cells with frequent mitotic figures specific to washings.

Of the non-epithelial elements, erythrocytes, leukocytes, and salt crystals are found. In the work of A.V. Zhuravleva points out that washouts should normally not contain formed elements. During inflammation, there is an increase in the number of cellular elements due to cells of the intermediate and basal zones, transitional epithelium and cells of an inflammatory nature.

In smears of a typical papilloma, cells of the same size are found: cylindrical, oval, round and spindle-shaped. “Tailed” cells may occur. Their nuclei are monomorphic with a compact chromatin structure and contain 1-2 nucleoli. The nuclear-cytoplasmic ratio is not disturbed.

According to G.A. Arzumanyan, characteristic is the absence of multinucleated cells characteristic of the upper, differentiated layer of the transitional epithelium. M.P. Ptokhov considers the most characteristic feature to be the presence of elongated caudate cells.

Cytological criteria for malignancy of papillomas

Cytograms of proliferating papillomas are characterized by more pronounced polymorphism and polychromasy of cells. The nuclei are increased in size, and the number of nucleoli in them increases. V.A. Enochovich finds the presence of oval and round tumor cells characteristic of this group of papillomas. N. Hanschke notes a large number of binucleate cells.

L.Ya. Lomonosov identifies the following cytological criteria for malignancy of papillomas:

Increased mitotic activity of cells;
polymorphism of nuclei with coarsening of the structure;
decrease in glycogen content in the cytoplasm;
an increase in cell layers of more than five.

He points out that sometimes incipient malignancy is easier to detect in cytograms than in histological preparations.

In transitional cell carcinoma of the bladder, all authors note polymorphism, polychromasia of cytogram cells, a violation of the nuclear-cytoplasmic ratio in favor of the nucleus, and changes in chromatin structure. hypertrophy and hyperplasia of the nucleoli and dystrophic changes in the cytoplasm. B.L. Polonsky and G.A. Arzumanyan considers multinucleation of cells characteristic of cancer.

Naturally, such a study must be carried out by an experienced specialist and requires a relatively long time. Therefore, in order to turn it into an automatic screening method, they began to use pulsed cytophotometry, which allows one to count 1000 cells stained with fluorescent dye in one second.

A number of works (Klein F. et al., 1982; Frankfurt O. et al., 1984; Dean P. et al., 1986) indicate a promising role in the diagnostic relationship of the state of cytological examination of urine and the flow cytometry method in which the measurement is made DNA, RNA content and nuclear size.

Pulse cytophotometry was used to quantitatively study DNA and proteins in epithelial cells of freely flowing urine, lavage specimens of the bladder, and suspensions from tumor tissue. Thus, a connection was identified between the presence of aneuploid cells in bladder lavages and the development of invasions in patients.

It has been established that most superficial bladder tumors are diploid, and invasion is accompanied by aneuploidy. Therefore, flow cytometric analysis of DNA content is a quantitative measure for predicting the degree of malignancy of a tumor, as well as for diagnosing cancer in situ.

In the work of K. Nielsen, a stereological assessment of the volume of the nuclei of the ileum of the bladder in normal and malignant processes is given. The author examined 27 biopsies from the bladder: 10 - normal, 5 - with infection, 12 - with tumors. A morphometric study established the average nuclear volume in normal conditions and during infection to be 133 and 182 µm3, and for cancer in situ - 536 µm3.

The test offered by the company "CYTODIAGNOSTICA", based on the quantitative determination of fluorescence, can help identify cells with an increased DNA content. The dye used binds cells with a high DNA content, and this circumstance makes it possible to identify one cancer cell among thousands of normal ones. The company proposes to use this test for screening populations at increased risk of bladder cancer.

But it should be noted that the use of the proposed methods for the prevention of bladder tumors encounters great difficulties, which begin immediately, already during urine collection - the cells in stale urine change significantly. It is clear that for preventive examinations the most acceptable cytological examination may be the examination of freshly released urine sediment; neither the aspiration biopsy method nor the method of washing from the mucous membrane of the bladder can become widespread.

The cytological examination of urine sediment, which we used to identify early oncological pathology of the bladder in the prophylactically examined contingents, did not live up to our hopes - not even once during the examination of 696 practically healthy individuals, of which 185 (38.6%) belonged to the risk group, and 511 (17.4%) - to the control, no signs of cell atypia were established, although the study was carried out by experienced cytologists in the central cytological laboratory of the city

Selected laboratory research methods

Certain laboratory research methods can identify a known predisposition to the occurrence of this form of cancer. We mean endogenous risk factors - impaired tryptophan metabolism with the appearance in the urine of its carcinogenic metabolites - 3-hydroxyanthranilic acid, 3-hydroxykynurenine, kynurenine, etc., as well as increased activity of urine β-hyaluronidase.

The presence of carcinogenic tryptophan metabolites in the urine indicates an increased risk of the disease; it is often determined in patients with bladder cancer, including recurrent ones, and indicates the need to correct tryptophan metabolism, i.e. engage in biochemical tumor prevention.

When forming risk groups using epidemiological tests, identifying among them those with the presence of carcinogenic tryptophan metabolites in the urine helps to further narrow the circle of people. requiring close attention from a urologist.

To determine the spectrum of tryptophan metabolites in the urine of subjects, one-dimensional descending paper chromatography can be used. Extraction of metabolites is carried out once from 100 ml of morning urine using the Makino method modified by Wachstein, Lobel.

To do this, urine is saturated with ammonium sulphide to precipitate proteins and salts. The urine is then filtered. 20 ml of molten phenol is added to the resulting filtrate and shaken for 20-25 minutes on a universal shaking apparatus. The resulting mixture is settled in a separating funnel, where two layers are quickly separated: the upper - yellow with a brown tint - phenol layer, the lower - colorless - urine layer.

The lower one is removed, 20 ml of sulfuric ether is added to the upper one and shaken vigorously to improve the dissolution of phenol in ether. In this case, phenol passes into the upper, lighter layer, and the ether-insoluble aqueous fraction containing aromatic derivatives of tryptophan remains in the lower part of the separatory funnel in the form of a dark oily layer with a volume of 0.5-0.7 ml.

This layer is carefully poured into an evaporation dish and dried in a fume hood. The dried extract is dissolved in 0.2 ml of distilled water and applied in an amount of 0.02 ml to chromatographic paper (Leningrad Slow), previously washed with a mixture of ether and alcohol in a ratio of 3:1.

After drying, the paper is placed in a chamber pre-saturated with solvents. The following solvent system is used as a solvent: N-butyl alcohol: glacial acetic acid: water - 4:1:1. Acceleration is carried out at room temperature for 20-24 hours.

After this, the chromatograms are dried and treated with Ehrlich's reagent (12 g of paradimethylaminobenzaldehyde + 20 ml of 6NHCI + 80 ml of ethyl alcohol). Immediately after this, yellow-orange spots appear on the chromatograms, which are produced with this reagent by urea and aromatic derivatives of tryptophan, and bluish-lilac spots caused by indole derivatives of the latter.

Based on the characteristic staining with Ehrlich's reagent, the Rf value and comparison with standards, the described method makes it possible to identify the following metabolites of tryptophan - urea, tryptamine, indican, tryptophan, 3-hydroxykynurenine, kynuranine, 3-hydroxyanthranilic acid. The latter is the most powerful endogenous carcinogen for the uroepithelium, gives a characteristic pinkish-orange staining with Ehrlich's reagent and an Rf value of 0.75-0.8.

Determination of urine kyaluronidase activity is carried out for the same purpose. But this method makes it possible to differentiate bladder papillomas from invasive cancer and is regarded by most authors as auxiliary in the diagnosis of epithelial tumors of the bladder

Enzyme activity in urine is determined by the generally accepted phenolphthalein method. To do this, urine is collected during the day under a thin layer of toluene, which protects it from decomposition; then its total volume, specific gravity, the presence of protein, leukocytes, sugar are determined, and microflora culture is necessarily carried out.

The latter is necessary due to the fact that various microorganisms, especially Escherichia coli, can be a source of β-hyaluronidase. In cases of bladder infection, urine is excluded from further testing.

Then 4-5 ml of urine is taken from each sample, which is centrifuged on a TsNL-2 centrifuge at 8-9 thousand revolutions for 8-10 minutes. Subsequently, according to the number of samples being studied, 0.5 ml of 0.1 M acite buffer is poured into graduated glass centrifuge tubes with a volume of 10 ml, 0.5 ml of a 0.05% solution of the substrate femolphthalein glucuronide (Sigma) is added, and only at the very end of the test and pre-centrifuged urine.

The addition of alkali made the pH of the mixture equal to 10.2-10.4, i.e., the optimum at which the most intense crimson-red coloration of the phenolphthalein separated under the influence of urinary β-hyaluronidase occurs. Further determination is carried out on an SF-4 spectrophotometer at a wavelength of 540 mm

The stained samples are compared with so-called "empty" baths, which contained the same solutions and in the same concentrations and quantities, but were not incubated and were prepared immediately before the determination of phenolphthalein in each test sample.

The amount of released phenolphthalein is determined using a pre-compiled calibration scale, followed by recalculation per 1 ml per 1 hour of incubation.

The level of enzyme activity when analyzing the urine of healthy individuals ranged from 0.4 to 1.1 and averaged 1.0 units. Fishman at 1 ml/hour. With papillomas of the bladder, there is an increase in enzyme activity by 1.2-1.4 times. Moreover, this increase is constant and does not depend on the ongoing treatment measures. In bladder cancer, β-hyaluronidase activity in urine is increased by no less than 2-3, and sometimes more times, reaching 5 Fishman units.

When determining the activity of this enzyme, one should take into account, firstly, concomitant diseases in patients. This applies, first of all, to hepatitis and pancreatitis, which themselves give a sharp increase in its activity in the urine. Secondly, it should be emphasized that even cystoscopy alone, not to mention more complex surgical interventions, immediately leads to an increase in the level of enzyme activity in the urine compared to the initial data, and only after 8-10 days the activity of β-lucuronidase returns to the original numbers.

According to M.N. Vlasova et al., which we present here, do not note any differences in the increase or decrease in enzyme activity depending on the age and gender of the patients, as well as chemotherapy or radiation treatment methods.

The authors use this method as an auxiliary method for diagnosing epithelial tumors in the bladder. It can also be recommended when carrying out measures for biochemical prevention of bladder cancer, aimed at reducing the activity of β-lucuronidase in the urine to control this activity.

Naturally, due to labor intensity and complexity, the method cannot be used as a screening method, but in risk groups its use is fully justified in order to identify individuals who have an important endogenous risk factor and, in this regard, need appropriate preventive measures.

Pryanichnikova M.B.

Cytological examination of urine, or analysis of urine for atypical cells, is a study of the structure of the elements of this biological fluid under a microscope. The material is assessed to determine the presence or absence of signs of malignant degeneration and other pathological processes in the cells. The method allows timely detection or control of diseases of the urinary system.

Purpose of the study

Urine is submitted for cytology in the following conditions:

suspicion of a neoplasm in the bladder, as well as the kidneys, ureters, urethra, prostate gland (prostate). The indication is hematuria - the presence of blood cells - red blood cells - in urine. In some cases, analysis is prescribed for urinary problems;
control of possible recurrence of urinary tract cancer;
the inability to use colposcopy and other methods to study the state of the reproductive system in women (virgins, during menstruation, with extensive inflammation). In this case, urine cells are examined.

Laboratory cytological examination is carried out to diagnose genitourinary cancer.

Urine sediment cytology cannot diagnose benign bladder tumors such as lipoma, fibroma, leiomyoma, neurofibromatosis, as well as pathological tissue proliferation - endometriosis. However, the method allows you to identify papillomas in time, preventing their degeneration, as well as detect cancer cells.

How to prepare for a cytological urine test

The study does not require special preparation. Before the procedure, it is advisable to perform genital hygiene and then collect the urine in a sterile container with a hermetically sealed lid.

It is advisable to deliver the sample to the laboratory within two hours.

The time for fluid collection depends on the direction of the study:

To analyze hormonal changes throughout a woman's menstrual cycle, the first portion of morning urine, which contains the most cellular elements, is required, although sometimes material taken at other times of the day is used.
To detect atypical cells, on the contrary, it is not recommended to study morning urine; it is better to wait 3 hours after the first urination and urinate in a container, collecting all the excreted liquid.
The most accurate results for bladder tumors are obtained by cytology of urine isolated by aspiration through a catheter - when the liquid is sucked out with a syringe from the organ cavity.

This method assesses human microflora

What will a cytological test tell you?

For atypical cells

Diagnosis of neoplasms of the urinary tract is based on the desquamation of the cells of these tumors and their release into urine:

In a benign process, individual cells or entire layers of transitional epithelium (the layer lining the inner surface of the bladder), the structure of which resembles the normal epithelium of the organ, are identified in the material. The shape of these elements is often spindle-shaped, together with them the presence of red blood cells is recorded in the urine.
In bladder cancer, transitional epithelial cells have pronounced signs of atypia - their structure is even different from each other. At the same time, the sample contains large numbers of red blood cells and necrotic masses.

In the laboratory, native (unmodified) smears and stained with special methods are prepared from the material sediment. Then the morphological composition of the cells is studied under a microscope. In addition to the presence of a benign or malignant tumor, cytological analysis helps to detect other lesions of the urinary tract, for example, an inflammatory process.

In addition, cytology is performed for those patients who are at the final stage of treatment of the disease.

The results of a study for atypical cells may be as follows:

Unsatisfactory sample - the collected urine is not suitable for testing (contains an insufficient number of cells or impurities that should not be in the material). The diagnosis needs to be repeated.
The test is negative - there are no cancer cells in the urine.
Atypical urine cytology - some changes were found in the cells of the sample, but without malignant signs.
Suspicious cytology – cellular material is not normal, possible cancer.
A positive test means there are malignant tumor cells in the urine.

The sensitivity of the method is about 90%. However, the study may have errors; this is affected by infectious lesions of the urinary tract, insufficient number of cells, stones in the bladder or kidneys, intravesical instillations (infusion of medications). If the test is positive, to confirm the diagnosis, they resort to cystoscopy - a biopsy (pinching off) of bladder tissue, followed by microscopic examination.

The advantage of this analysis is that the cytological test does not require much time compared to other studies

For hormonal fluctuations

In this case, native and stained smears are also studied, and then the number of cells of different types in them is counted, including:

basal;
intermediate;
basophilic (keratinizing);
acidophilic (keratinized);
non-nuclear acidophilus.

Particular attention is paid to the latter - the number of these elements on different days of the cycle is 2-20% and indicates the release of hormones from the adrenal glands (more than from the ovaries).

Other indicators

Microscopy of urine sediment is part of a general urine analysis and can reveal other pathologies. For example, when the level of leukocytes is higher than normal, they speak of an inflammatory process, and the detection of bacteria or fungi indicates urinary tract infections.

Cytological examination of urine allows us to identify tumor lesions of the urinary system, as well as non-tumor changes in the epithelium, and is an additional method for the early diagnosis of urothelial cancer, including high-grade cancer (HGC). The non-invasiveness and accessibility of the cytological method allows monitoring of patients with a history of cancer in order to detect relapses of the disease.

SynonymsEnglish

Cytological examination.

Research method

Cytological method.

What biomaterial can be used for research?

Daily urine.

How to properly prepare for research?

Eliminate alcohol from your diet for 24 hours before the test.
Avoid (in consultation with your doctor) taking diuretics for 48 hours before collecting urine.

General information about the study

Urine is essentially a biological product of the breakdown and elimination of metabolic processes in the body; it can be used to judge both the state of the body as a whole and the presence of certain diseases, processes and pathologies.

There are several types of urine laboratory tests. Biochemical, clinical, daily, urine analysis according to Nechiporenko, Zimnitsky, Reberg, Sulkovich test, bacteriological culture - each method has certain indications and features of its implementation. Conventionally, they can be divided into specific and nonspecific. The above methods are among the first, and nonspecific methods - immunochromatographic and chemical-toxicological - make it possible to determine the presence and percentage of alcohol-containing substances.

Cytological examination of urine, or analysis of urine for atypical cells, is a study of the structure of urine elements under a microscope. The biomaterial is assessed to determine the presence or absence of signs of malignant degeneration and other pathological processes in the cells. The analysis makes it possible to assess the characteristics of the morphological structure of individual cells. Thanks to this, the doctor can subsequently confirm or refute the primary diagnosis, and, if necessary, prescribe additional medical examination. This is one of the most effective ways to detect malignant tumors and cancer. The study also helps in identifying infectious and inflammatory processes.

Cytological examination differs in that it is carried out on cellular level. A study of cell morphology using a microscope is carried out in order to identify malignant tumors or oncological formations. The state of the human microflora is assessed, the presence or absence of pathological infectious and other processes is indicated, the following types of diseases can be determined: prostate cancer, bladder cancer, cancer of the ureters and urethra, kidney cancer. The method is considered one of the most effective ways to detect cancerous degenerations at the initial stage.

What is the research used for?

To assess the characteristics of the morphological structure of cellular elements in urine, to identify diseases of the genitourinary system.

When is the study scheduled?

If you suspect the presence of urothelial cancer;
identifying a group of patients at high risk of developing VZZ cancer for further examination;
examination of patients with benign processes (urolithiasis, cystitis, benign prostatic hypertrophy, kidney disease, diabetes mellitus, inverted papilloma, hyperplasia, nephrogenic adenoma);
clarification of the diagnosis for existing oncology of the urinary tract (bladder carcinoma);
hematuria;
assessment of treatment effectiveness;
monitoring the results of the operation.

What do the results mean?

The cytological report is a description of the general cytological picture.

The result must be interpreted in accordance with clinical signs and history.

Patients with a history of VZZ urothelial carcinoma and no clinical signs of tumor may have an undiagnosed tumor in the upper urinary tract. For such patients, detection in cytospecimens atypical urothelial cells (AUC) means a high probability of clinical detection of a high-grade tumor.

If atypical cells are detected and/or characteristic clinical symptoms indicating the presence of a malignant neoplasm, it is recommended to carry out further targeted laboratory and instrumental diagnostics (cystoscopy, biopsy).

General urine analysis with sediment microscopy

Determination of the risk of occurrence and unfavorable course of urothelial bladder cancer, determination of p16ink4a in urine sediment

Bladder Cancer Antigen (UBC)

Who orders the study?