Clinical and Experimental Morphology

06/10/2021

Recurrent catamenial tension pneumothorax in a patient with endometriosis

Free full text available: http://cem-journal.ru/index.php/cem/article/view/135 (In Russian)

G.A. Kazaryan1, D.V. Bazarov1, A.Yu. Grigorchuk1, A.A. Kazaryan2, V. Yu. Savina1, S.A. Levakov2, S.V. Paukov2, A.E. Mamedova2, G.Ya. Azadova2

1 B.V. Petrovsky National Research Centre of Surgery, Moscow, Russia
2 I.M. Sechenov First Moscow State Medical University of the Ministry of Health of Russia (Sechenov University), Moscow, Russia

DOI: https://doi.org/10.31088/CEM2021.10.2.57-61

Abstract
Catamenial pneumothorax is a rare cause of spontaneous pneumothorax in women of reproductive age and one of the thoracic endometriosis manifestations. Researchers have shown an increased interest in catamenial pneumothorax but a clear treatment strategy for this pathology has not been developed yet. The thoracoscopic revision of the pleural cavity, as well as the identification and subsequent histological verification of ectopic endometriosis foci, play a crucial role in patient management. This case report describes the successful treatment of a patient with recurrent catamenial tension pneumothorax and following multidisciplinary analysis.

Keywords: catamenial pneumothorax, thoracic endometriosis, histological examination

05/10/2021

Protocol optimization for obtaining the culture of rat dermal fibroblasts

Free full text available: http://cem-journal.ru/index.php/cem/article/view/136 (In Russian)

E.A. Ponomarenko, M.A. Diatroptova, K.A. Artemyeva, A.Yu. Shelkov
Research Institute of Human Morphology, Moscow, Russia

DOI: https://doi.org/10.31088/CEM2021.10.2.62-69

Abstract
Introduction. Obtaining primary cultures of fibroblasts is necessary for conducting experimental studies to investigate the basic cell response mechanisms to various stimuli. Despite the widespread use of fibroblast cultures, methods for obtaining them from the skin are not standardized. The aim of the study was to find the optimal conditions to obtain a fibroblast culture from rat skin using enzymatic tissue disaggregation.

Materials and methods. The fibroblast culture was obtained from the dermis of 18 male Wistar rats (N=12, 5–6-week-old, and 60–65 g body weight; N=6, 8–10-week-old rats, and 160–180 g body weight). A series of experiments was carried out to obtain a fibroblast culture with selecting the collagenase type and defining its concentration and exposure time.

Results. An optimized protocol to obtain rat dermis fibroblasts is presented. Moreover, the problems of obtaining a culture and practical aspects of its use are discussed.

Conclusion. When obtaining a primary culture of rat skin fibroblasts we should consider the following factors: the type of the enzyme, its concentration, and exposure time; the age of the animals; the area of skin graft collection. In young rats, the optimal result was achieved when the cells were isolated from the axillary zone using collagenase type II at a 1 mg/ml concentration during a 90-minute exposure. In adult animals, the enzymatic effect of collagenase type II on the skin graft obtained from the back was optimal at a 5 mg/ml concentration during a 120-minute exposure.

Keywords: fibroblast culture, dermis, preparation protocols, rats

04/10/2021

Structural dermis remodeling in a skin expansion rat model

Free full text available from: http://cem-journal.ru/index.php/cem/article/view/134 (in Russian)

E.S. Mishina1, M.A. Zatolokina1, M.V. Mnikhovich2

1 Kursk State Medical University, Kursk, Russia
2 Research Institute of Human Morphology, Moscow, Russia

DOI: https://doi.org/10.31088/CEM2021.10.2.50-56

Abstract
Introduction. Skin expansion is known to be the most effective way to obtain skin for alloplastic closing of large defects, and it has proven to be successfully used in various fields of surgery, cosmetology, and traumatology. At the same time, the issues of skin flap restructuring when it is stretched and possible range of its application are still relevant. In this regard, the aim was to study the adaptive skin rearrangements of the fibrous skeleton in a murine skin expansion model.

Materials and methods. We used the skin of mature male Wistar rats (N=30) after a 2-week stretching. We utilized a complex morphological light and electron microscopy approach as well as an immunohistochemical method to determine the types of collagen and to study the alterations in skin samples.

Results. We revealed pronounced reactive changes in the skin structural components in the skin expansion area. On day 14, we observed a decrease in the epidermal cell layer thickness as well as stretching, partial disorganization, and damage of dermal fibrous structure. At the same time, cellular mechanisms of regeneration and activation of collagen synthesis were launched, the morphological substrate of which was (1) a significant 2.7-fold increase in the number density of fibroblasts, which produce substances for building fibrous structures, (2) a five-fold increase in macrophages, which are a catalyst for the fibroblast functional activity and (3) a collagen fiber types’ redistribution s with Type III collagen structures’ predominance.

Conclusion. The results indicate, first of all, the restructuring of the dermal fibrous component, which implies reparative and restorative processes. These must be taken into account in clinical practice to achieve not only an esthetic effect but also the subsequent adequate functioning and vital activity of the skin flap.

Keywords: skin expansion, fibrous dermis, remodeling, skin, hyperextension

03/10/2021

Morphological features of spermatogenesis in rats after targeted irradiation with electrons of variable doses

Free full text available from: http://cem-journal.ru/index.php/cem/article/view/133 (in Russian)

G.A. Demyashkin1,2

1 National Medical Research Radiological Centre of the Ministry of Health of Russia, Moscow, Russia
2 I.M. Sechenov First Moscow State Medical University of the Ministry of Health of Russia (Sechenov University), Moscow, Russia

DOI: https://doi.org/10.31088/CEM2021.10.2.40-49

Abstract
Introduction. Today, a primary concern of humanity is male infertility. Recent figures suggest that the disease affects 186 million people worldwide, and its prevalence increases as men get older. In literature the radiation therapy effect on the male reproductive system is debated. Over the past decade, scientific and technological progress has definitely accelerated the development of the radiation therapy methods, and as new devices have emerged, there is a need to carefully select new dosing regimens. The aim of the experimental study was to evaluate morphological spermatogenesis indicators after targeted irradiation with electrons of variable doses.

Materials and methods. Wistar rats (n=70) were once subjected to targeted ionizing irradiation on a pulsed electron accelerator NOVAC 11 with doses of 2 Gy, 4 Gy, 6 Gy, 8 Gy, 10 Gy, and 12 Gy, and a week later spermatogenesis was assessed with light microscopy.

Results. In the experimental groups, we revealed a decrease in the number of germ cells; signs of degenerative changes and lysis of spermatids and spermatozoa; the appearance of intense pyknotic nuclei in primary spermatocytes; and a sharp decrease in the number of spermatogonia and Sertoli and Leydig cells without change in their structure. In the irradiation groups of 8–12 Gy, there was the deepest damage to the tubules, up to spermatogenic epithelium aplasia, and extensive vacuolization.

Conclusion. In the te**es, we observed dose-dependent progressive pathological changes in tissue structure as well as a decrease in the number of spermatogonia and other germ cells on day 7 after irradiation with a pulsed electron accelerator NOVAC 11.

Keywords: electron irradiation, male infertility, spermatogenesis, seminal globs

02/10/2021

PU.1 is a nuclear factor of immunocompetent cells of tumor stroma in colorectal cancer

Free full text available from: http://cem-journal.ru/index.php/cem/article/view/132 (in Russian)

O.V. Kovaleva1, A.N. Gratchev1, P.A. Podlesnaya1, M.A. Rashidova1, D.V. Samoilova1, N.Yu. Sokolov1, Z.Z. Mamedli1, D.A. Kudlay1, N.E. Kushlinskii1

1 N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Health of Russia, Moscow, Russia
2 I.M. Sechenov First Moscow State Medical University of the Ministry of Health of Russia (Sechenov University), Moscow, Russia

DOI: https://doi.org/10.31088/CEM2021.10.2.32-39

Abstract
Introduction. Tumor-associated macrophages (TAMs) are traditionally considered to be a pro-tumor factor that promotes the growth of various tumors; however, for colorectal carcinomas (CRC), the prognostic significance of TAMs has not been fully determined, which may be due to the lack of macrophage markers suitable for this tumor type. The aim of this work was to study the expression of the nuclear marker of stromal cells PU.1 in colorectal tumors and its association with the clinical and morphological tumor characteristics.

Materials and methods. We performed an immunohistochemical analysis to assess the expression of PU.1, CD68, and CD20 in 85 primary CRCs. The Mann-Whitney test was used to determine statistically significant differences in independent groups. Correlation analysis of the expression of the studied protein was carried out by determining the Spearman’s rank correlation coefficient. Differences were considered statistically significant at p

01/10/2021

Extragenital noninfectious pathology in the maternal mortality structure in Moscow

Free full text available from: http://cem-journal.ru/index.php/cem/article/view/131 (in Russian)

L.M. Mikhaleva1,5, A.G. Konoplyannikov2,6, Y.Y. Kudryavtseva3, A.S. Olenev4, N.A. Gracheva5, A.E. Biryukov1,5, S.A. Mikhalev6, E.N. Songolova7

1 Research Institute of Human Morphology, Moscow, Russia
2 Pirogov Russian National Research Medical University, Moscow, Russia
3 Peoples’ Friendship University of Russia, Moscow, Russia
4 Perinatal Center of the City Clinical Hospital No. 24 of the Moscow Healthcare Department, Moscow, Russia
5 City Clinical Hospital No. 31 of the Moscow Healthcare Department, Moscow, Russia
6 Family Planning and Reproduction Center of the Moscow Healthcare Department, Moscow, Russia
7 L.A. Vorokhobov City Clinical Hospital No. 67 of the Moscow Healthcare Department, Moscow, Russia

DOI: https://doi.org/10.31088/CEM2021.10.2.21-31

Abstract
Introduction. Diseases directly related to pregnancy are still leading the maternеal mortality structure, although in many countries there is an increase in the number of deaths from extragenital pathology among pregnant women. A significant number of lethal cases in this group are associated with environmental factors, lifestyle, and the overall increase in somatic diseases. The aim of the study was to determine the main causes of maternal mortality and investigate the role of extragenital pathology based on autopsy data.

Materials and methods. The study included 29 autopsy observations. We studied medical records, autopsy protocols of deceased pregnant women, women in labor, and puerperant women. Each cause of death was classified according to ICD-10. A clinical and morphological study was also carried out using additional histochemical and immunohistochemical staining methods.

Results. The data from pathological studies show that in Moscow, extragenital pathology accounts for up to 40% of maternal deaths. The leading cause of death is circulatory system diseases, including an arteriovenous malformation; a myocardial infarction; a congenital heart disease (patent foramen ovale); cerebral, aortic, and splenic artery aneurysms. The second most common one is other specified diseases and conditions complicating pregnancy. This group of diseases included malignant neoplasms. Less common causes of death are gastrointestinal diseases, other diseases of the blood and blood-forming organs and certain disorders involving the immune mechanism that complicate pregnancy, and also respiratory diseases. A single case is associated with a generalized viral infection.

Conclusion. Extragenital pathology remains an important obstetric problem, and the management of pregnant women with systemic diseases requires a multidisciplinary treatment approach.

Keywords: maternal mortality, extragenital pathology, arteriovenous malformation, cerebral aneurysm, antiphospholipid syndrome

30/09/2021

Epithelial-mesenchymal transition in main types of gastric carcinoma

Free full text available: http://cem-journal.ru/index.php/cem/article/view/130 (in English)

I.V. Vasilenko, R.B. Kondratyk, I.S. Grekov and A.M. Yarkov

M. Gorky Donetsk National Medical University, Donetsk

DOI: https://doi.org/10.31088/CEM2021.10.2.13-20

Abstract
Introduction. The rapid development of basic science enabled us to significantly expand our understanding of various intercellular interactions. Epithelial-mesenchymal transition (EMT) is known to play a key role in certain tissue formation in the embryonic period. However, recent data show that EMT can also be observed in some pathological conditions, in particular, in various neoplasm development. This suggests that there are a number of alternative and fundamentally new mechanisms for the tumor formation and progression. Thus, EMT, which occurs in carcinomas, increases the invasiveness, immunoresistance, immunity to therapy, and the metastatic potential. Knowledge of EMT features and their timely recognition in morphological tumor diagnosis is of great predictive importance for patients. The aim of the research was to study the morphological features of epithelial-mesenchymal transition in the main types of gastric cancer.

Materials and methods. We studied specimens of gastric carcinomas (N=64) including 31 cases of diffuse type, 19 cases of intestinal type, and 14 cases of mixed type.

Results. All cases of the diffuse carcinoma group showed spread EMT features, which appeared already in the mucosa and completed with positive vimentin expression in 93.5% of cases. The malignant cell proliferative activity was low; however, in 29% of cases we detected areas of moderate or even high activity. In the intestinal type gastric cancer, EMT developed as a result of tumor progression, it arose more often in the deeper layers and was incomplete and focal. As a rule, the proliferative activity of tumor cells was high and moderate. Vascular invasion occurred more often in diffuse type (90.3%), less often in mixed type (71.4%), and even less often in the intestine type (55.8%) gastric carcinoma.

Conclusion. The variety of morphological features of EMT, its frequency, prevalence, completeness, and sequence in the development of various types of gastric cancer determines the features of their clinical manifestation and influences their further management.

Keywords: gastric cancer, diagnosis, histological main types, EMT, morphopathology

29/09/2021

Cryopreservation of tissue-engineered constructs in regenerative medicine

Free full text: http://cem-journal.ru/index.php/cem/article/view/129 (In Russian)

I.V. Arutyunyan, T.K. Dubovaya

1 V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology of Ministry of Health of Russia, Moscow, Russia
2 Pirogov Russian National Research Medical University, Moscow, Russia

DOI: https://doi.org/10.31088/CEM2021.10.2.6-12
Abstract

The transplantation of artificial tissues and organs is gradually becoming a part of our reality. At the same time, researchers are facing a problem common to all transplantologists, i.e. the need for a long-term storage of a biomedical product (transplant) without losing its properties. The possibility to cryopreserve cells adhered to various scaffolds' surface was first presented about 20 years ago. However, the data on the technology as a whole remains unsystematized and controversial. This review aimed to analyze the literature on tissue-engineered constructs (TEC) cryopreservation of different scientific groups to create a unified approach in assessing the technique's efficacy necessary for further regenerative medicine development. The comparison of studies on TEC cryopreservation conducted by various research groups is hampered not only by the lack of standardized protocols but also by different approaches to assessing the result. As experimental data were accumulated, the cryopreservation efficacy was reassessed from meeting the basic requirements for the structure preservation (thawed TEC retains its integrity, cells are partially alive and attached to the matrix) to focusing on the final result (thawed TEC retains its functional properties and is ready to be transplanted). Many of the currently used in vitro research methods presented in the review allow one to look for new ways of increasing the TEC cryopreservation efficacy; however, in our opinion, the next step on the way to introducing the technology into clinical practice should be researched on experimental animals.

Keywords: tissue engineered construction, cryopreservation, efficacy estimation

29/09/2021

№2 2021 Volume 10 online
Full text available:

электронный журнал: Клиническая и экспериментальная морфология

06/07/2021

Muir–Torre syndrome case associated with a hereditary MSH2 mutation

Free full text available: http://cem-journal.ru/index.php/cem/article/view/126 (In Russian)

A.V. Kaminskiy1, I.L. Plaksa2,3

1 I.P. Pavlov Ryazan State Medical University, Ryazan, Russia
2 Leningrad Regional Clinical Oncological Dispensary, Saint Petersburg, Russia
3 GENETICO LLC, Moscow, Russia

DOI: 10.31088/CEM2021.10.1.66-70

Abstract
Muir–Torre syndrome combines skin tumors with sebaceous differentiation with malignant tumors of the gastrointestinal tract, which is caused by hereditary mutations in the genes of the mismatched nucleotide repair system. The article presents a clinical case of a 42-year old man who complained of an exophytic skin tumor on the back measuring 2.3 × 1.8 cm. At histological examination, the tumor was presented primarily with sebaceous differentiatied cells, which corresponded to adenoma of the sebaceous glands. From the patient’s history it became known that at the age of 37 he underwent hemicolectomy for high-differentiated adenocarcinoma. An immunohistochemical study with antibodies to proteins of the mismatched nucleotide repair system in a tumor sample revealed the absence of a nuclear reaction in tumor cells with antibodies to MLH1 and MSH2 proteins, which indicates the presence of germline mutations in these genes. High-throughput semiconductor parallel DNA sequencing revealed a variant of the nucleotide sequence in exon 12 of the MSH2 gene (c.1797_1801 del), leading to a shift in the reading frame (NM_000251: p.L599fs).

Keywords: Muir–Torre syndrome, colorectal cancer, repair system, wrong-paired bases

06/07/2021

Experimental model of chronic focal osteomyelitis for in vivo studies

Free full text available: http://cem-journal.ru/index.php/cem/article/view/127 (In Russian)

V.N. Mitrofanov, O.P. Zhivtsov, N.Yu. Orlinskaya, D.V. Davydenko

Privolzhsky Research Medical University of the Ministry of Health of Russia, Nizhny Novgorod, Russia

DOI: 10.31088/CEM2021.10.1.71-77

Abstract
Introduction. The article describes an experimental model of chronic suppurative osteomyelitis in a rabbit. As new therapeutic and diagnostic algorithms for the supervision of patients with osteomyelitis appear, there is an growing need to compare the methods of surgical debridement and plasty of bone defects in an infectious process, in order to create experimental standardized pathological conditions as close to the clinical course of the disease in humans as possible. The aim of the study was to develop an experimental model of a standardized chronic purulent bone cavity, suitable for a comprehensive assessment of surgical debridement effectiveness and osteoplastic properties of bone substitute materials.

Materials and methods. A standardized defect of the tibia in 24 rabbits was formed. The Staphylococcus aureus strain was used as an infectious agent. A dynamic assessment of the main indicators of blood counts in animals was carried out. The formation of chronic osteomyelitis was evaluated using radiography, computed tomography methods and histological studies.

Results. It was shown that purulent bone wound developed in experimental animals with the technique created, and a defect with signs of a chronic purulent-inflammatory process was demonstrated.

Conclusion. The proposed model of chronic osteomyelitis is reproducible. Operational flexibility and identical in size and location bone defects allow to use this model in new osteoplastic material research.

Keywords: chronic osteomyelitis, experiment, experimental animals

06/07/2021

Comparative morphological and biochemical characteristics of the toxic effects of doxorubicin and nanosomal PLGA-doxorubicin form in the experimental glioblastoma treatment

Free full text available from: http://cem-journal.ru/index.php/cem/article/view/125 (in Russian)

V.V. Kudelkina1, A.S. Khalanskiy1, O.V. Makarova1, I.S. Tsvetkov1, A.M. Kosyreva1, A.I. Alekseeva1, A.Y. Shelkоv1, O.О. Maksimenko2, V.A. Razzhivina2, S.E. Gelperina2,3

1 Research Institute of Human Morphology, Moscow, Russia
2 LLC Drugs Technology, Khimki, Russia
3 D.I. Mendeleev University of Chemical Technology of Russia, Moscow, Russia

DOI: 10.31088/CEM2021.10.1.58-65

Abstract
Introduction. Doxorubicin (Dox) in the composition of poly(lactic-co-glycolic acid, 50:50) (PLGA) – nanoparticles has high antitumor efficacy in rats with glioblastoma 101.8. However, the toxic effect of Dox-PLGA is not well understood. The aim of the study was morphological and biochemical evaluation of the hepatotoxic and cardiotoxic effects of doxorubicin and Dox-PLGA in the glioblastoma 101.8 treatment in Wistar rats.

Materials and methods. The study was performed on 24 male Wistar rats with glioblastoma: no treatment (n=7), treated with doxorubicin (n=9) or Dox-PLGA (n=8) intravenously at a dose of 1.5 mg/kg on days 2, 5 and 8 after tumor implantation. On the 14th day of the experiment, morphological changes in the myocardium and liver were examined. Hematological and biochemical blood tests were performed.

Results.When treating rats with experimental glioblastoma 101.8, Dox-PLGA in comparison with doxorubicin has less pronounced cardiotoxic and hepatotoxic effects according to the morphological, hematological and biochemical tests. Inflammatory changes in the myocardium of the animals treated with Dox-PLGA were less pronounced and widespread than the ones treated with doxorubicin. The activities of total and cardiac creatine phosphokinase (CPK) isoforms and AST were statistically significantly lower in Dox-PLGA group than in animals with glioblastoma without treatment and receiving doxorubicin. The hepatotoxic effects of Dox-PLGA were minimal. Unlike animals treated with doxorubicin, they had mild hepatocyte dystrophy. ALT activity in all groups did not differ from the reference values.

Conclusion. Compared with doxorubicin, the nanosomal form of Dox-PLGA in the experimental glioblastoma 101.8 treatment has less pronounced cardio- and hepatotoxic effects.

Keywords: rat glioblastoma 101.8, doxorubicin, PLGA-nanoparticles, toxic effect, liver, myocardium, morphology, biochemistry