Intercellular Substance Research Articles

Essential oils cause membrane disruption and autoaggregation of MDR Acinetobacter baumannii cells

Essential oils are promising antimicrobial agents against various bacteria. The aim of the study was to examine anti-Acinetobacter baumannii activity of nine essential oils and to elucidate essential oil mechanisms of action against this. In total, 8 out of 9 examined different essential oils exhibited activity against multidrug-resistant (MDR) A. baumannii isolates. The best anti-A. baumannii activity expressed Thymus serpyllum, Satureja hortensis and Oreganum majorana essential oil (MBC 1.29 to 2.58 µg mL−1), while essential oils of Menta x piperita, Hyssopus officinalis, Foeniculum vulgare and Artemisia dracunculus expressed the same effect in higher concentrations (1.80 to 3.80 µg mL−1). The effect of Juniperus sabina, Juniperus sibirica and S. hortensis essential oils against A. baumannii is for the first time reported here. The mechanisms of Myrtus communis, Eucalyptus camaldulensis, S. hortensis and T. serpyllum essential oils were further examined. The essential oil treatments led to concentration dependent leakage of biomolecules form the cells, causing an increase in the concentrations of lipids, carbohydrates, and proteins in the suspensions. Microstructural observations confirmed the essential oil effect on cell membranes and disruption the membrane integrity, with obvious leakage of intercellular substance, collapsed cells with perforations, debris presence and autoaggregated cells. The subinhibitory concentrations of oils did not obviously changed protein patterns determined by SDS-PAGE. These results indicate that the essential oils, particularly T. serpyllum, S. hortensis, M. communis and E. camaldulensis express its anti-A. baumannii activity via membrane disruption and increased membrane permeability, representing very promising alternative antibacterial agents against MDR wound isolates.

Renal Mast Cell-Specific Proteases in the Pathogenesis of Tubulointerstitial Fibrosis.

Chronic kidney disease is detected in 8-15% of the world's population. Along with fibrotic changes, it can lead to a complete loss of organ function. Therefore, a better understanding of the onset of the pathological process is required. To address this issue, we examined the interaction between mast cells (MCs) and cells in fibrous and intact regions, focusing on the role of MC proteases such as tryptase, chymase, and carboxypeptidase A3 (CPA3). MCs appear to be involved in the development of inflammatory and fibrotic changes through the targeted secretion of tryptase, chymase, and CPA3 to the vascular endothelium, nephron epithelium, interstitial cells, and components of intercellular substances. Protease-based phenotyping of renal MCs showed that tryptase-positive MCs were the most common phenotype at all anatomic sites. The infiltration of MC in different anatomic sites of the kidney with an associated release of protease content was accompanied by a loss of contact between the epithelium and the basement membrane, indicating the active participation of MCs in the formation and development of fibrogenic niches in the kidney. These findings may contribute to the development of novel strategies for the treatment of tubulointerstitial fibrosis.

The therapeutic use of exosomes in children with adenoid hypertrophy accompanied by otitis media with effusion (AHOME): a protocol study

BackgroundThe adenoids act as a reservoir of bacterial pathogens and immune molecules, and they are significantly involved in children with otitis media with effusion (OME). As an essential carrier of intercellular substance transfer and signal transduction, exosomes with different biological functions can be secreted by various types of cells. There remains significant uncertainty regarding the clinical relevance of exosomes to OME, especially in its pathophysiologic development. In this study, we will seek to determine the biological functions of exosomes in children with adenoid hypertrophy accompanied by OME (AHOME).MethodsThe diagnostic criteria for OME in children aged 4–10 years include a disease duration of at least 3 months, type B or C acoustic immittance, and varying degrees of conductive hearing loss. Adenoidal hypertrophy is diagnosed when nasal endoscopy shows at least 60% adenoidal occlusion in the nostrils or when nasopharyngeal lateral X-ray shows A/N > 0.6. Children who meet the indications for adenoidectomy surgery undergo adenoidectomy. Peripheral blood, nasopharyngeal swab, and adenoid tissue will be collected from patients, and the exosomes will be isolated from the samples. Following the initial collection, patients will undergo adenoidectomy and peripheral blood and nasopharyngeal swabs will be collected again after 3 months.Expected resultsThis study aims to identify differences in exosomes from preoperative adenoid tissue and peripheral blood samples between children with AHOME and those with adenoid hypertrophy alone. Additionally, it seeks to determine changes in microbial diversity in adenoid tissue between these groups.ConclusionsThe findings are expected to provide new insights into the diagnosis and treatment of OME, to identify novel biomarkers, and to enhance our understanding of the pathophysiology of OME, potentially leading to the development of innovative diagnostic and therapeutic approaches.

Inhibition of Prolactin Affects Epididymal Morphology by Decreasing the Secretion of Estradiol in Cashmere Bucks.

Yanshan Cashmere bucks are seasonal breeding animals and an important national genetic resource. This study aimed to investigate the involvement of prolactin (PRL) in the epididymal function of bucks. Twenty eleven-month-old Cashmere bucks were randomly divided into a control (CON) group and a bromocriptine (BCR, a prolactin inhibitor, 0.06 mg/kg body weight (BW)) treatment group. The experiment was conducted from September to October 2020 in Qinhuangdao City, China, and lasted for 30 days. Blood was collected on the last day before the BCR treatment (day 0) and on the 15th and 30th days after the BCR treatment (days 15 and 30). On the 30th day, all bucks were transported to the local slaughterhouse, where epididymal samples were collected immediately after slaughter. The left epididymis was preserved in 4% paraformaldehyde for histological observation, and the right epididymis was immediately preserved in liquid nitrogen for RNA sequencing (RNA-seq). The results show that the PRL inhibitor reduced the serum PRL and estradiol (E2) concentrations (p < 0.05) and tended to decrease luteinizing hormone (LH) concentrations (p = 0.052) by the 30th day, but no differences (p > 0.05) occurred by either day 0 or 15. There were no differences (p > 0.05) observed in the follicle-stimulating hormone (FSH), testosterone (T), and dihydrotestosterone (DHT) concentrations between the two groups. The PRL receptor (PRLR) protein was mainly located in the cytoplasm and intercellular substance of the epididymal epithelial cells. The PRL inhibitor decreased (p < 0.05) the expression of the PRLR protein in the epididymis. In the BCR group, the height of the epididymal epithelium in the caput and cauda increased, as did the diameter of the epididymal duct in the caput (p < 0.05). However, the diameter of the cauda epididymal duct decreased (p < 0.05). Thereafter, a total of 358 differentially expressed genes (DEGs) were identified in the epididymal tissues, among which 191 were upregulated and 167 were downregulated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that ESR2, MAPK10, JUN, ACTL7A, and CALML4 were mainly enriched in the estrogen signaling pathway, steroid binding, calcium ion binding, the GnRH signaling pathway, the cAMP signaling pathway, and the chemical carcinogenesis-reactive oxygen species pathway, which are related to epididymal function. In conclusion, the inhibition of PRL may affect the structure of the epididymis by reducing the expression of the PRLR protein and the secretion of E2. ESR2, MAPK10, JUN, ACTL7A, and CALML4 could be the key genes of PRL in its regulation of epididymal reproductive function.

Retinal pigment epithelial cell culture

Objective: Retinal pigment epithelial cells (RPEs), as the main cellular component of the preretinal membrane, whose excessive proliferation can lead to the development of proliferative vitreoretinopathy as interaction with other cellular components and intercellular substance. Here, we developed an effective method to culture RPEs so as to provide effective cell source for later experimental usage. Methods: The harvested of eye cup that retains the retinal pigment epithelium was gently washed three times with phosphate buffered saline (PBS), and was added with 0.02 mg/ml dispase enzyme, which was cut into small pieces and then placed in the incubator for digestion for about half an hour. Subsequently, gently blowing was performed multiple times to detach the RPEs from the retina to collect the RPEs suspension. After implantation into the culture dish, when the cells were grown and covered with 80-90% of the bottom of the bottle, digestion of the cells is carried out with 0.25% trypsin. After removing them from the wall, the digested cell suspension was collected, followed by addition of Dulbecco's modified Eagle medium/Nutrient Mixture F-12 (DMEM/F12) to stop digestion. The mixture was subject to centrifuge at 1000 r/min for 8 min, with the supernatant discarded, and then the cells located in bottom were passaged at 1:3 ratios. During the process, the morphology and viability were observed and cell identification were also performed. Results: Primary cultured cells began to adhere to the wall at 4 h, characterized with a flat and polygonal morphology, rich in pigment particles. On 3 days, there was a significant increase in cell numbers accompanied by cluster formation, resulting in a stabilized cobblestone-like appearance. By 5 days, cells elongated and formed dense clusters. Growth curve analysis revealed robust proliferation at 3 and 5 days. Immunofluorescence staining confirmed that over 90% of the cells were positive for RPE-65, indicating successful isolation and culture of pure rat RPEs. Conclusions: The RPEs cultured by the above method are in good growth and it can be used for further experimental studies.

High-sensitivity THz-ATR imaging of cerebral ischemia in a rat model.

The fast label-free detection of the extent and degree of cerebral ischemia has been the difficulty and hotspot for precise and accurate neurosurgery. We experimentally demonstrated that the fresh cerebral tissues at different ischemic stages within 24 hours can be well distinguished from the normal tissues using terahertz (THz) attenuated total reflection (ATR) imaging system. It was indicated that the total reflectivity of THz wave for ischemic cerebral tissues was lower than that for normal tissues. Especially, compared to the images stained with 2,3,5-triphenyl tetrazolium chloride (TTC), the ischemic tissues can be detected using THz wave with high sensitivity as early as the ischemic time of 2.5 hours, where THz images showed the ischemic areas became larger and diffused as the ischemic time increasing. Furthermore, the THz spectroscopy of cerebral ischemic tissues at different ischemic times was obtained in the range of 0.5-2.0 THz. The absorption coefficient of ischemic tissue increased with the increase of ischemic time, whereas the refractive index decreased with prolonging the ischemic time. Additionally, it was found from hematoxylin and eosin (H&E) staining microscopic images that, with the ischemic time increasing, the cell size and cell density of the ischemic tissues decreased, whereas the intercellular substance of the ischemic tissues increased. The result showed that THz recognition mechanism of the ischemia is mainly based on the increase of intercellular substance, especially water content, which has a stronger impact on absorption of THz wave than that of cell density. Thus, THz imaging has great potential for recognition of cerebral ischemia and it may become a new method for intraoperative real-time guidance, recognition in situ, and precise excision.

The importance of polymorphism of the COL1A1 gene in the development of pelvic organ prolapse in women of childbearing age

Pelvic organ prolapse (POP) is a chronic slowly progressive disease. One of the most significant risk factors for the formation of omissions and prolapses of the internal genitalia is connective tissue diseases. Type I collagen is the most common protein of the intercellular substance of connective tissue, mutation of the COL1A1 gene encoding it leads to the synthesis of a defective protein. The aim of our study was to investigate the significance of COL1A1 gene polymorphism in POP development and in the occurrence of relapses of the disease in operated patients of different age groups. Material and methods. The study included 68 patients with verified pelvic organ prolapse, divided into two age groups (n = 34 in each): the first group included women aged from 23 to 44 years, the second – aged from 46 to 72 years. In all patients, the presence of manifestations of undifferentiated connective tissue dysplasia (UCTD) was assessed, a laboratory study of the polymorphism gene COL1A1 was conducted and the content of type 1 C-terminal collagen peptide (β-CrossLaps) in venous blood was measures. Results and discussion. 86.8 % of the examined patients had clinical signs of UCTD. Statistical analysis indicates that there is a relationship of the frequency of UCTD detection and severity of manifestations with age: the age of the patients with moderate to heavy UCTD (38 [37; 41] years, median [lower quartile; upper quartile]) was statistically significantly lower than that of the women with light degree (45.5 [38; 62] years; p &lt; 0.001 years) and without UCTD (56 [48; 65] years; p &lt; 0.001). There were no statistically significant differences in the polymorphism of the COL1A1 gene between patients under 45 (1 group) and over 45 (2 group) (p = 0.25), however, there is a tendency to decrease the frequency of G/G and increase T/T types of polymorphism in women of reproductive age. In patients with recurrent POP after surgical treatment, G/G polymorphism was less common (p &lt; 0.001) and G/T was more common (p = 0.04). The level of β-CrossLaps was higher than the normative level in 8.8 % of the examined women; in 2 cases of identified heterozygous (G/T) polymorphisms, its significant rise was noted. ROC analysis performed to identify the relationship between age, gene COL1A1 polymorphism, signs of UCTD and the level of β-CrossLaps indicates the effectiveness of the integrated use of these indicators as predictors of POP development in women of the reproductive period. Conclusions. The results of the study suggest that the features of gene COL1A1 polymorphism, the level of β-CrossLaps can serve as predictors of the development of POP in women of reproductive age with clinical manifestations of signs of UCTD. Complex of these indicators allows to develop a prognostic model of POP early manifestation.

Effect of Liquid Ammonia on the Structure and Properties of Superfine Wool Fibers

Superfine wool fiber has a soft and luxurious texture, excellent moisture absorption and is warmer and more comfortable than ordinary wool. Liquid ammonia, which has exceptional permeability, exerts a certain modification effect on the surface scales of wool in the finishing and processing of wool fabric, thereby enhancing its wear performance. However, limited research has been conducted on the impact of liquid ammonia on valuable wool fibers such as superfine wool and cashmere. In this article, superfine wool tops were treated with continuous liquid ammonia finishing equipment and the effects of liquid ammonia on the microstructure and surface morphology of superfine wool fiber were investigated. In addition, the changes in length, fineness, anti-felting, and dyeing properties of superfine wool fiber after liquid ammonia treatment were examined. The results demonstrated that the interstitial integrity of intercellular substance of superfine wool was compromised following treatment with liquid ammonia, which had a greater tendency to infiltrate cortical cells and induce alterations in wool structure and performance. Furthermore, the surface scales of superfine wool were damaged after treatment with liquid ammonia, leading to improved anti-felting and dyeing properties.

Exosomes: efficient macrophage-related immunomodulators in chronic lung diseases.

Macrophages, the predominant immune cells in the lungs, play a pivotal role in maintaining the delicate balance of the pulmonary immune microenvironment. However, in chronic inflammatory lung diseases and lung cancer, macrophage phenotypes undergo distinct transitions, with M1-predominant macrophages promoting inflammatory damage and M2-predominant macrophages fostering cancer progression. Exosomes, as critical mediators of intercellular signaling and substance exchange, participate in pathological reshaping of macrophages during development of pulmonary inflammatory diseases and lung cancer. Specifically, in inflammatory lung diseases, exosomes promote the pro-inflammatory phenotype of macrophages, suppress the anti-inflammatory phenotype, and subsequently, exosomes released by reshaped macrophages further exacerbate inflammatory damage. In cancer, exosomes promote pro-tumor tumor-associated macrophages (TAMs); inhibit anti-tumor TAMs; and exosomes released by TAMs further enhance tumor proliferation, metastasis, and resistance to chemotherapy. Simultaneously, exosomes exhibit a dual role, holding the potential to transmit immune-modulating molecules and load therapeutic agents and offering prospects for restoring immune dysregulation in macrophages during chronic inflammatory lung diseases and lung cancer. In chronic inflammatory lung diseases, this is manifested by exosomes reshaping anti-inflammatory macrophages, inhibiting pro-inflammatory macrophages, and alleviating inflammatory damage post-reshaping. In lung cancer, exosomes reshape anti-tumor macrophages, inhibit pro-tumor macrophages, and reshaped macrophages secrete exosomes that suppress lung cancer development. Looking ahead, efficient and targeted exosome-based therapies may emerge as a promising direction for treatment of pulmonary diseases.

Cell-in-cell-mediated intercellular communication exacerbates the pro-inflammatory progression in asthma.

Cell-in-cell (CIC) structures have been suggested to mediate intracellular substance transport between cells and have been found widely in inflammatory lung tissue of asthma. The aim of this study was to investigate the significance of CIC structures in inflammatory progress of asthma. CIC structures and related inflammatory pathways were analyzed in asthmatic lung tissue and normal lung tissue of mouse model. In vitro, the activation of inflammatory pathways by CIC-mediated intercellular communication was analyzed by RNA-Seq and verified by Western blotting and immunofluorescence. Results showed that CIC structures of lymphocytes and alveolar epithelial cells in asthmatic lung tissue mediated intercellular substance (such as mitochondria) transfer and promoted pro-inflammation in two phases. At early phase, internal lymphocytes triggered inflammasome-dependent pro-inflammation and cell death of itself. Then, degraded lymphocytes released cellular contents such as mitochondria inside alveolar epithelial cells, further activated multi-pattern-recognition receptors and NF-kappa B signaling pathways of alveolar epithelial cells, and thereby amplified pro-inflammatory response in asthma. Our work supplements the mechanism of asthma pro-inflammation progression from the perspective of CIC structure of lymphocytes and alveolar epithelial cells, and provides a new idea for anti-inflammatory therapy of asthma.

Preparation of Nanoporous Carbon from Rice Husk with Improved Textural Characteristics for Hydrogen Sorption

This study proposes a method to control the pore-forming process by performing preliminary mechanical activation of the initial rice husk before carbonization. Preliminary mechanical activation of the initial rice husk leads to the loosening of the intercellular substance and its partial depolymerization, thereby increasing the availability of its internal structure for pore formation during carbonization and chemical activation. Using the method described above, nanoporous carbon was obtained with a Brunauer–Emmett–Teller (BET)-calculated specific surface area of 2713 m2/g, a micropore specific surface area calculated by using the Dubinina–Radushkevich (D-R) method of 3099 m2/g, and a total pore volume calculated by using the Barett–Joyner–Halenda (BJH) method of 1.625 cm3/g. Due to these characteristics, the adsorption capacity in the obtained sample was for hydrogen 3.7 wt.% at a temperature of −190 °C and a pressure of 9 kgf/cm2, which is 29.7% higher than the adsorption capacity of nanoporous carbon obtained based on rice husk without mechanical activation. The composite “carbon–platinum” NC-2/Pt10%, at a temperature of 20 °C and a pressure of 9 kgf/cm2, showed an increase in sorption capacity of 27% compared to pure nanoporous carbon NC-2, which is explained by the emergence of the spillover effect.

Diagnostic and Therapeutic Roles of Extracellular Vesicles and Their Enwrapped ncRNAs in Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is a systemic inflammatory disease whose precise pathogenesis remains mysterious. The involvement of epigenetic regulation in the pathogenesis of RA is one of the most anticipated findings, among which non-coding RNAs (ncRNAs) hold great application promise as diagnostic and therapeutic biomarkers for RA. Extracellular vesicles (EVs) are a heterogeneous group of nano-sized, membrane-enclosed vesicles that mediate intercellular communication and substance exchange, especially the transfer of ncRNAs from donor cells, thereby regulating the functional activities and biological processes of recipient cells. In light of the significant correlation between EVs, ncRNAs, and RA, we first documented expression levels of EVs and their-encapsulated ncRNAs in RA individuals, and methodically discussed their-implicated signaling pathways and phenotypic changes. The last but not least, we paied special attention to the therapeutic benefits of gene therapy reagents specifically imitating or silencing candidate ncRNAs with exosomes as carriers on RA animal models, and briefly highlighted their clinical application advantage and foreground. In conclusion, the present review may be conducive to a deeper comprehension of the diagnostic and therapeutic roles of EVs-enwrapped ncRNAs in RA, with special emphasis on exosomal ncRNAs, which may offer hints for the monitoring and treatment of RA.

Panax notoginseng saponins reverse steroid resistance in lupus nephritis: Involvement of the suppression of exosomal P-gp levels from lymphocytes to glomerular endothelial cells

Microangiopathy is the most basic pathological manifestation of lupus nephritis (LN), and glomerular endothelial cells (GECs) injury is an important pathological mechanism. LN patients with microangiopathy are prone to steroid resistance (SR). Our previous studies confirmed that Panax notoginseng saponins (PNS) could reverse SR by downregulating the expression of P-gp in SR lymphocytes of LN mice (SLCsL/S). However, the mechanism of how circulating lymphocytes transmit SR information to GECs and thus affect the efficacy of kidney treatment is not clear. Recent studies have found that exosomes (exos) are an important carrier for intercellular bioactive substance communication. But whether exosomes derived from SLCsL/S mediate SR in GECs and PNS interventions. To solve this problem, Exosomes isolated from SLCsL/S were characterized, and in vitro cell coculture was further conducted to investigate the effect of SLCsL/S-derived exosomes in the SR of GECs and PNS intervention. Sequencing was used to define the exosomal miRNA expression profiling of SR GECs. Moreover, the in vivo experiments were performed through the injection of exosomes extracted from SLCsL/S into the tail vein of mice.Our research results indicate that exosomes derived from SLCsL/S could transmit SR information to GECs and lead to the aggravation of inflammatory injury through conferring P-gp, which were negated by a P-gp inhibitor. Further, we identified higher levels of exosomal miR-125b-5p from SR GECs were associated with SR in LN and could serve as biomarker for the risk of developing SR. PNS could reverse the SR of GECs and alleviate inflammatory injury by suppressing exosomal P-gp levels from lymphocytes to GECs in vitro and in vivo. However, the specific molecular mechanism by which PNS regulates exosomes has not yet been elucidated, and we need to conduct more in-depth research in the future. Overall, Our findings suggest that exosomal transfer of SLCsL/S derived P-gp confer SR to GECs, and PNS can target exosome communication to reverse SR in LN, which provides new ideas and a scientific basis for improving the clinical efficacy of traditional Chinese medicine in the treatment of refractory LN.

WntA and Wnt4 during the regeneration of internal organs in the holothurian Eupentacta fraudatrix.

Over the past few years, it has been established that wnt genes are involved in the regenerative processes of holothurians. The wnt4 gene was identified as one of the most active genes in Eupentacta fraudatrix regeneration using differential gene expression analysis and qPCR of individual genes. Also, the wntA gene was found in holothurians, which is present only in invertebrates and can perform unique functions. In this regard, both these genes and proteins were studied in this work. During regeneration, the Wnt4 protein is found in the cells of the coelomic and ambulacral epithelium, retractor muscles, and radial nerves. Single cells with this protein are also found in the connective tissue of the developing aquapharyngeal bulb and in the hypoderm of the body wall. Cells with WntA are found exclusively in the hypoderm of the body wall. We assume that both genes are involved in regeneration, but Wnt4 coordinates the formation of the epithelial tissue structure, while WntA maintains the state of the intercellular substance of the body wall.

Гидролизованный морской коллаген в комплексной терапии дегенеративно-дистрофических изменений опорно-двигательного аппарата

Degenerative-dystrophic changes in the musculoskeletal system of various localizations are one of the most common reasons for patients to turn not only to a rheumatologist, but also to a neurologist and first-contact doctors (therapist and general practitioner) who provide primary health care. Medicines that modify the symptoms of degenerative-dystrophic changes in the musculoskeletal system are divided into “fast-acting” (analgesics, NSAIDs, tramadol) and “slow-acting” (chondroprotectors). When considering “slow-acting” drugs, many publications are devoted to the natural components of the cartilage intercellular substance – glucosamine and chondroitin sulfate, while much less attention is paid to the importance of collagen. In articular cartilage, collagens provide most of the biomechanical properties of the extracellular matrix necessary for its function. Collagen hydrolysates have demonstrated positive results in clinical trials related to joint health, such as reducing joint pain, increasing mobility, and improving joint structure. Marine collagen is the closest in molecular structure to human collagen fibers. Its main characteristics include high biological activity and biocompatibility, as well as low antigenicity. The possible effects of vitamin C, vitamin D3, zinc and calcium in the complex treatment of degenerative-dystrophic changes in the musculoskeletal system are considered. A biological supplement containing vitamins, microelements and hydrolyzed marine collagen Promerim® optimizes the prevention and treatment of patients with degenerative changes in the musculoskeletal system.

MORPHOLOGICAL AND MORPHOMETRIC CHANGES OF CHONDROCYTES OF THE JOINT SURFACE OF THE PATELLA IN THE CONDITIONS OF EXPERIMENTAL DIABETIC KNEE ARTHROPATHY

Diabetes is a widespread disease that has a tendency to grow and often leads to disability and mortality of the population at a young working age. In this pathology, the joints are often aff ected. According to many researchers, more than 50 % of patients with type 1 and type 2 diabetes are diagnosed with arthropathies, i. e. diabetes- associated osteoarthritis. Features of structural changes of chondrocytes of the articular surface of the patella in diabetic arthropathy of the knee joint have not been sufficiently investigated. The purpose of the study was to study the features of structural changes in the chondrocytes of the articular surface of the patella in experimental diabetic arthropathy of the knee joint. The chondrocytes of the articular surface of the patella of 60 laboratory sexually mature white male rats, which were divided into 3 groups, were investigated using a complex of morphological methods. The 1-st group included 20 intact practically healthy animals, the 2-nd – 20 rats with a one-month experimental diabetic arthropathy of the knee joint, the 3-rd – 20 experimental animals with the two-month specifi ed pathology. Diabetes mellitus was modeled by a single intraperitoneal injection of streptozotocin from the company «Sigma» in a dose of 50 mg/kg. One month and two months after the start of the experiment, rats were euthanized by bloodletting under thiopental anesthesia. The diameters of young and mature chondrocytes and their nuclei, the nuclear- cytoplasmic ratio in these cells, the relative volume of damaged chondrocytes, and intercellular- chondrocyte ratios were determined on histological microsections in the cartilage tissue of the articular surface of the patella. Quantitative indicators were processed statistically. It was established that long-term hyperglycemia leads to diabetic arthropathy of the knee joint, which is characterized by atrophy of chondrocytes of the articular surface of the patella and changes in their nuclear- cytoplasmic ratios, which indicated a violation of cellular structural homeostasis. It was found that the nuclear- cytoplasmic ratio in young chondrocytes of the articular surface of the patella was changed by 5.3 % (р&lt;0.01) during one-month experimental hyperglycemia, by 8.4 % (р&lt;0.001) during two-month hyperglycemia, in mature chondrocytes – by 3.8 % (p&lt;0.05) and 7.7 % (p&lt;0.001). In diabetic arthropathy, a marked increase in the relative volume of damaged chondrocytes and the volume of intercellular substance was noted. The detected changes in quantitative morphological indicators depended on the duration of hyperglycemia and dominated in young chondrocytes.

High level of tumor marker CA19-9 returned to normal after cholecystectomy in calculous cholecystitis patients

Background High serum CA19-9 is usually caused by pancreaticobiliary malignancies, but it has also been found in a tiny minority of calculous cholecystitis patients. Aims To clarify the relationship between calculous cholecystitis and serum CA19-9. Methods Clinical data of calculous cholecystitis patients with high serum CA19-9 (high group, n = 20) and normal serum CA19-9 (normal group, n = 40) who underwent cholecystectomy were analyzed. Serum CA19-9 of high group were followed-up and gallbladder specimens were analyzed by immunohistochemistry. Results Serum CA19-9 in the high group ranged from 105 to 1635 U/ml, of which 30% exceeded 1000 U/ml. Follow-up results showed that 20 patient’s serum CA19-9 returned to normal after cholecystectomy, including 4 closely followed-up patients whose serum CA19-9 recovered within one month. Immunohistochemical results revealed that CA19-9 was mildly positive only in mucosal epithelial cells in the normal group, but positive in mucosal epithelial cells, vascular endothelial cells, and intercellular substances in the high group, accounting for high serum CA19-9. Conclusion Serum CA19-9 is proved to be associated with calculous cholecystitis for the first time, so that clinicians should consider calculous cholecystitis associated CA19-9 elevation in the clinic practice besides other CA19-9 related diseases.

Pre-Sacral Chordomas, about Two Cases and Review of the Literature

The World Health Organization (WHO) defines chordoma as a malignant tumor of intermediate or low grade, developing at the expense of embryonic remnants of the notochord. These tumors represent 8.4% of all malignant bone tumors. We report 2 cases collected in the medical oncology department at the CHU Hassan II in Fes. In whom the clinical examination revealed a mass and/or signs of loco-regional compression. The tumor was located in the sacrum in both patients. The diagnosis was based on the data of the anatomopathological examination showing the association of a lobulated architecture with classic physaliphore cells and a mucoid intercellular substance. Immunohistochemistry was performed for both patients. The principle of therapeutic management was based on radiotherapy and targeted therapy since surgery was difficult given the local tumor extension.

THERAPY OF PERIODONTITIS WITH VASCULAR ENDOTHELIAL GROWTH FACTOR BASED ON HYDROGEL: IMMUNOHISTOCHEMICAL CHARACTERISTICS

Regeneration of lost periodontal tissues is the ultimate goal of periodontal treatment. One of the promising approaches to stimulating periodontal regeneration is the use of hydrogels and various growth factors that contribute to the colonization of the scaffold with fibroblast populations, followed by the formation of mature connective tissue of the periodontal ligament. Patients with chronic generalized periodontitis of mild and moderate severity (n=70, age 49-69 years) were treated in the Department of maxillofacial surgery of the UKB No. 4. Treatment was carried out with a preparation from a complex of bioresorbable carbohydrates and recombinant human vascular endothelial growth factor (VEGF-A). The biopsy material was taken immediately before the introduction of the drug into the periodontal pocket and 14 days after the procedure. Prior to treatment, pronounced chronic inflammation with a predominance of lymphocytes, moderate edema and divergence of the fibers of the stromal component were noted in the gum preparations, signs of chronic moderate active inflammation were observed. After treatment, collagen fibers constructed from type III collagen and type I collagen were found in the structured fibers of the intercellular substance, which is characteristic of the formed connective tissue. Thus, the therapy of chronic periodontitis with a hydrogel with VEGF-A is a promising direction for further research, since it has the potential to influence not only stimulating the regeneration of periodontal tissues, but also the effect of local anti-inflammatory treatment.

Functions and therapeutic potentials of exosomes in osteosarcoma.

Osteosarcoma is a primary malignant tumor of the skeleton with the morbidity of 2.5 in 1 million. The regularly on-set is in the epiphysis of the extremities with a high possibility of early metastasis, rapid progression, and poor prognosis. The survival rate of patients with metastatic or recurrent osteosarcoma remains low, and novel diagnostic and therapeutic methods are urgently needed. Exosomes are extracellular vesicles 30-150 nm in diameter secreted by various cells that are widely present in various body fluids. Exosomes are abundant in biologically active components such as proteins, nucleic acids, and lipids. Exosomes participate in numerous physiological and pathological processes via intercellular substance exchange and signaling. This review presents the novel findings of exosomes in osteosarcoma in diagnosis, prognosis, and therapeutic aspects.