The use of postmortem (autopsy) material in fundamental and applied biomedical research significantly facilitates the collection of biomaterial for statistically robust sample cohorts. However, natural adaptive processes to developing cellular stress in the early postmortem period, caused by oxygen and nutrient deprivation, trigger the activation of numerous genes promoting cell survival under stress. Many of these activated pathways are also crucial for tumor cell survival in vivo, as evidenced by various transcriptomic studies. This study aimed to investigate the potential influence of postmortem interval (PMI) duration on gene expression in normal and tumor tissues. Using a model of chemically induced hepatocellular carcinoma in mouse liver, we comparatively analyzed the dynamics of transcript levels for several genes (BRCA1, BRCA2, CHEK1, CHEK2, ATM, CDK12) in paired samples of normal and tumor tissue over a 24-h PMI using RT-qPCR. In normal tissue, gene expression increased significantly, while tumor tissue demonstrated relative transcriptional stability, with no substantial changes in the studied transcript levels. A critical finding was the observed convergence of expression profiles: initial differences between the tissues were completely eliminated by 24 h PMI. This pattern developed despite formally adequate RNA quality (RQN) and the absence of clear signs of progressive autolysis in histology, indicating the insufficiency of standard quality criteria for detecting postmortem changes. These findings collectively underscore the critical importance of minimizing and controlling PMI during the biobanking of oncological samples for reliable transcriptomic research.

The review provides a basic description of extracellular matrix proteins and their corresponding receptors expressed by tumor cells that are involved in key mechanisms of metastasis. According to the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines, 479 of the initially identified 8,112 publications met the inclusion criteria (212 in English and 267 in Russian). The final analysis included 38 articles relevant to the topic of this review, three of which were in Russian. The presented data expand the fundamental knowledge about the molecular mechanisms of metastasis and can be used as a basis for the development of targeted anticancer compounds.

Iron deficiency anemia (IDA) remains a global health challenge. This study pioneers the use of humic substances (HS) as natural, biocompatible macroligands to develop safer and more effective nanoferrotherapeutics. We synthesized a series of nanoscale Fe(III) oxyhydroxide complexes stabilized by different HS, employing various solvents (ethanol, isopropanol, and acetone) and precipitation methods to isolate fractions with optimized properties. The nanocomposites were comprehensively characterized using inductively coupled plasma atomic emission spectrometry, total organic carbon analysis, X-ray diffraction, transmission electron microscopy, and Mössbauer spectroscopy. Cytotoxicity and iron bioavailability of all HS-Fe(III) formulations were assessed in Caco-2 intestinal epithelial cells. The type of HS and precipitation conditions significantly influenced the nanocomposites’ properties, yielding spherical nanoparticles (1–2 nm) of ferrihydrite or goethite. Physicochemical analysis confirmed that solvent-driven fractionation effectively tailored the nanocomposites’ size, crystallinity, and elemental composition. All HS-Fe(III) formulations demonstrated exceptional cytocompatibility, starkly contrasting the significant cytotoxicity of the reference drug Ferrum Lek®. Several complexes, particularly CHSFe-Et67, surpassed Ferrum Lek® in cellular iron uptake efficiency. We conclude that HS are a highly promising platform for developing effective and safe iron-delivery nanoferrotherapeutics, leveraging their natural polyfunctionality to enhance bioavailability and mitigate toxicity.

Even though the survival and proliferation stages of cancer cells that have newly settled at a metastatic site are the rate-limiting stages and the most promising targets for drugs, there is a lack of models of the earliest stage of metastasis formation. A method for modeling breast cancer liver metastasis is described here: a stage of transition of a differentiated tumor cell into a cell actively proliferating in a three-dimensional (3D) liver spheroid. Opposite to existing heterocellular 3D models of metastases, the protocol allows modeling the initial stage of liver colonization by metastatic cells, the so-called “micrometastases.” The method includes obtaining a line of fluorescent tumor cells, fluorescence-activated sorting of differentiated cells, preparing a single-cell suspension of liver cells, forming a liver spheroid in an agarose mold, inducing the tumor cell dedifferentiation and proliferation using IL-6, and intravital microscopy of spheroids, with subsequent processing and analysis of fluorescent images in the ImageJ software. The performance of the proposed model was demonstrated using microRNA therapeutics. The ability of a combination of microRNAs to suppress the transition of micrometastasis to macrometastasis in the 3D liver spheroid was confirmed by an immunofluorescent assay of spheroid sections and transcriptome analysis.

Aim. To examine the pattern of morphological changes, RNA quality number, and gene expression in mouse tissues sampled at autopsy under controlled experimental conditions.Materials and methods. Balb/c mice were euthanized and subsequently subjected to necropsy at 0, 3, 12, 24, 48, and 72 hours of the postmortem period. During the first three hours following euthanasia, the mice were maintained at room temperature, after which they were transferred to a refrigerator (4 º С). Total RNA was extracted from tissue samples taken from the kidney, liver, and brain; the integrity of the RNA samples was assessed by capillary electrophoresis, and the RNA quality number (RQN) was calculated. The expression levels of Actb, Epas1, and Rps18 housekeeping genes were evaluated by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) with original primers and probes using the TaqMan assay. The histologic examination was performed according to standard techniques.Results. Degradation of RNA extracted from mouse kidney tissues appeared to be greater than that of RNA taken from the liver. In the meantime, a negative linear correlation was observed between RQN and the duration of the postmortem interval for liver and kidney samples. In contrast, no significant changes in the RQN score were observed for brain RNA samples at any of the time points. The expression of the Epas1 and Rps18 genes was significantly decreased in mouse kidney and liver tissues. However, the level of Epas1 and Rps18 gene expression in the brain remained stable at all time points and did not exhibit a significant decrease at 72 hours after euthanasia. No obvious morphological changes were detected by the histologic examination, which does not exclude the presence of ultrastructural pathological changes.Conclusion. RQN in autopsy tissues serves as a crucial predictor of sample quality for molecular biology studies, including gene expression analysis.

We propose an original strategy for metastasis prevention using a combination of three microRNAs that blocks the dedifferentiation of cancer cells in a metastatic niche owing to the downregulation of stemness genes. Transcriptome microarray analysis was applied to identify the effects of a mixture of microRNAs on the pattern of differentially expressed genes in human breast cancer cell lines. Treatment of differentiated CD44(-) cancer cells with the microRNA mixture inhibited their ability to form mammospheres in vitro. The combination of these three microRNAs encapsulated into lipid nanoparticles prevented lung metastasis in a mouse model of spontaneous metastasis. The mixture of three microRNAs (miR-195-5p/miR-520a/miR-630) holds promise for the development of an antimetastatic therapeutic that blocks tumor cell dedifferentiation, which occurs at secondary tumor sites and determines the transition of micrometastases to macrometastases.

Ischemic stroke (IS) is a prevalent condition worldwide with high mortality and disability risks. The issue of its pathogenetic therapy remains unresolved by now due to the limited effectiveness and safety of reperfusion measures. Recent research has elucidated that neuroinflammation plays a pivotal role in IS development and may serve as a therapeutic target. The NLRP3 inflammasome emerges as a key mediator orchestrating post-ischemic inflammatory reactions through activation of caspase-1, which cleaves pro-IL-1β and pro-IL-18 into active pro-inflammatory cytokines released into the extracellular milieu. This review presents insights into the structure and activation process of the NLRP3 inflammasome in IS. The factors and mechanisms contributing to both its activation and inhibition are outlined.

Due to traumatic injuries, including those from surgical procedures, adhesions occur in over 50% of cases, necessitating exclusive surgical intervention for treatment. However, preventive measures can be implemented during abdominal organ surgeries. These measures involve creating a barrier around internal organs to forestall adhesion formation in the postoperative phase. Yet, the effectiveness of the artificial barrier relies on considerations of its biocompatibility and the avoidance of adverse effects on the body. This study explores the biocompatibility aspects, encompassing hemocompatibility, cytotoxicity, and antibacterial and antioxidant activities, as well as the adhesion of blood serum proteins and macrophages to the surface of new composite film materials. The materials, derived from the sodium salt of carboxymethylcellulose modified by glycoluril and allantoin, were investigated. The research reveals that film materials with a heterocyclic fragment exhibit biocompatibility comparable to commercially used samples in surgery. Notably, film samples developed with glycoluril outperform the effects of commercial samples in certain aspects.

The features of the participation of Smad3 in the functioning of neural stem cells (NSC), neuronal committed precursors (NCP), and neuroglial elements were studied in vitro. It was found that this intracellular signaling molecule enhances the clonogenic and proliferative activities of NCP and inhibits specialization of neuronal precursors. At the same time, Smad3 does not participate in the realization of the growth potential of NSC. With regard to the secretory function (production of neurotrophic growth factors) of neuroglial cells, the stimulating role of Smad3-mediated signaling was shown. These results indicate the promise of studying the possibility of using Smad3 as a fundamentally new target for neuroregenerative agents.

An investigation was carried out on humic substances (HSs) isolated from the coal of the Kansk-Achinsk basin (Krasnoyarsk Territory, Russia). The coal HSs demonstrate the main parameters of molecular structure inherent to this class of natural compounds. An assessment was performed for the chemical, microbiological, and pharmacological safety parameters, as well as the biological efficacy. The HS sample meets the safety requirements in microbiological purity, toxic metals content (lead, cadmium, mercury, arsenic), and radionuclides. The presence of 11 essential elements was determined. The absence of general, systemic toxicity, cytotoxicity, and allergenic properties was demonstrated. The coal HS sample was classified as a Class V hazard (low danger substances). High antioxidant and antiradical activities and immunotropic and cytoprotective properties were identified. The ability of the HS to inhibit hydroxyl radicals and superoxide anion radicals was revealed. Pronounced actoprotective and nootropic activities were also demonstrated in vivo. Intragastric administration of the HS sample resulted in the improvement of physical parameters in mice as assessed by the “swim exhaustion” test. Furthermore, intragastric administration in mice with cholinergic dysfunction led to a higher ability of animals with scopolamine-induced amnesia to form conditioned reflexes. These findings suggest that the studied HS sample is a safe and effective natural substance, making it suitable for use as a dietary bioactive supplement.

The aim of this review was to analyze the accumulated data on the use of mass spectrometry in diagnosing, treating, and prognosing cancer from the perspective of precision medicine. Currently, universally accepted methods for early cancer diagnosis are not available, primarily due to low molecular specificity of pathological changes at early stages of cancer development. Additionally, the existing diagnostic modalities are notably limited in sensitivity. However, early detection is imperative for selection of the most suitable cancer treatment strategy and its successful implementation. In the realm of oncology, mass spectrometry approaches show great potential for advancement and utilization. Mass spectrometry is becoming an indispensable tool in basic and applied research due to its sensitivity, specificity, and accuracy. It allows for efficient analysis of complex biological compounds, even at low concentrations. Moreover, contemporary mass spectrometry technology is capable of automating the analysis, thereby facilitating its diverse clinical applications in diagnosis, drug therapy selection, and even potential assistance to surgical oncologists in the operating room. Considering all these characteristics and advantages, mass spectrometry methods for the analysis of biological samples can be defined as some of the most promising and dynamically developing tools in precision medicine, as they are capable of providing clinically valuable information based on omics technologies, taking into account personal characteristics of the patient. Over the next decade, introduction of mass spectrometry-based methods into clinical practice based on the principles of precision medicine is expected to optimize selection of personalized treatment strategies for cancer patients and provide significant economic benefits by reducing morbidity, disability, and mortality.This comprehensive review presents the analysis of 65 scientific publications, highlighting the results of clinical and experimental studies utilizing mass spectrometry methods for diagnosing cancer, investigating the underlying mechanisms of disease development, and evaluating the efficacy of therapeutic interventions. The review encompasses original articles published from January 1, 2018 to November 30, 2023. The majority of studies back the potential of mass spectrometry as a valuable tool for cancer diagnosis and treatment monitoring. Broadening application of mass spectrometry techniques in the field of oncology holds significant promise and represents a relevant area for future research.

Aim. To develop a new method to determine the viability of Opisthorchis felineus in vitro using the MTS reagent and to evaluate its applicability for analyzing the efficacy of anthelmintic agents in the treatment of opisthorchiasis.Materials and methods. Golden hamsters were used to create a model of O. felineus infection. The animals were infected with metacercariae obtained from fish of the Cyprinidae family. Three months after infection, adult parasites were extracted from the hepatobiliary system. Their viability was assessed using the motility scale and a new method based on the modified MTS test protocol. To account for differences between the size and number of adult parasite cells, the results were normalized with respect to protein content. To evaluate the feasibility of the new approach in the study of pharmacological activity against opisthorchiasis, the viability of adult parasites in the presence of praziquantel was tested.Results. During incubation of adult flukes in a medium with the addition of the MTS reagent, colored watersoluble formazan was accumulated. Thermal inactivation of parasites significantly decreased the production of this compound. Since the studied adult parasites differed in size and number of cells, the obtained data on their viability were normalized to protein content. The results correlated with the data on parasite viability obtained by the traditional method using the motility scale. Evaluation of praziquantel efficacy at different concentrations using two independent methods (the MTS test and the motility scale) showed that the results of the MTS test were consistent with literature data and comparable with the results obtained using the motility scale.Conclusion. A new method for in vitro evaluation of anti-opisthorchiasis activity of drugs was developed. It is based on the assessment of water-soluble formazan production by adult O. felineus flukes in the culture medium using the MTS reagent for screening anti-opisthorchiasis activity of new anthelmintic drugs.

In in vitro model of short-term therapeutic inhalation of Xe/O2 mixture, xenon in millimolar concentrations led to a pronounced decrease in induced platelet aggregation in the platelet-enriched blood plasma. The maximum and statistically significant decrease occurred in response to induction by collagen (by ≈30

Aim. To conduct a clinical and morphological assessment of the characteristics of colon cancer depending on the dMMR / pMMR status of the tumor. Materials and methods . A retrospective study included 66 patients with operable colorectal cancer (CRC) (T1-4bN0-2bM1), who were treated at Cancer Research Institute of Tomsk National Research Medical Center (NRMC). The average age of the patients was 64.4 ± 12.8 years. All patients underwent hemicolectomy or colon resection, as well as intraoperative resection of distant metastases, if present. Results. We determined that in CRC patients with pMMR tumors, hematogenous metastases were detected in 27.3% of cases, while in patients with dMMR tumors, hematogenous metastases were detected only in 6.1% of cases ( p = 0.021). A comparative analysis of dMMR and pMMR tumors also allowed to establish higher frequency of perineural invasion among the pMMR subgroup of carcinomas ( p = 0.039). The sign of tumor budding was found both in dMMR carcinomas (36%) and in pMMR tumors (45%). This sign was associated with damage to regional lymph nodes ( p = 0.0017). A more detailed analysis of the tumor budding phenomenon showed that in dMMR tumors, Bd1 low-grade budding (83%) predominated. In pMMR tumors, Bd2 intermediate-grade budding (33%) and Bd3 high-grade budding (26.7%) prevailed. Bd2 and Bd3 tumor budding types were associated with hematogenous metastasis ( p < 0.001). Conclusion. The obtained data demonstrate the differences in such pathomorphological parameters as perineural invasion and the degree of tumor budding depending on the dMMR / pMMR status of the tumor. These histologic parameters in tumor tissue are also associated with higher incidence of distant metastasis in patients with pMMR carcinomas as opposed to patients with dMMR tumors.

Experimental type 1 diabetes mellitus (T1DM) was induced in rats by daily intraperitoneal injections of alloxan in a dose of 90 mg/kg for 4 days. For verification of insulin resistance, insulin tolerance test was performed in 2 weeks and the glucose utilization rate constant (KITT) was calculated. The rats demonstrated the main symptoms of T1DM: hypoinsulinemia, hyperglycemia, ketonemia, glucosuria, ketonuria, polydipsia, polyphagia, weight loss, and insulin resistance, as evidenced by a decrease in KITT. The serum content of free fatty acids and triacylglycerols significantly increased. The content of triacylglycerols increased in skeletal muscles and decreased in the liver. A negative linear correlation was found between KITT and triacylglycerol content in muscles. Thus, the development of insulin resistance in experimental T1DM in rats is associated with accumulation of triacylglycerols in skeletal muscles.

The comparative phytochemical analysis in this study revealed differences in the type and levels of phenolic compounds between Spiraea hypericifolia L. and Spiraea crenata L. The compounds in water–ethanol extracts of aerial parts of both species were identified by high-performance liquid chromatography as chlorogenic, gentisic, and cinnamic acids; quercetin; kaempferol; hyperoside; isoquercetin; nicotiflorin; and apigenin. In the extract of S. hypericifolia, p-coumaric acid and luteolin were also found, which were absent in the extract of S. crenata. Such compounds as avicularin, astragalin, and isorhamnetin-3-rutinoside proved to be specific to S. crenata (and were not found in the S. hypericifolia extract). The viability of liver cancer HepG2 cells and breast cancer MDA-MB-231 cells significantly decreased after cultivation with the S. crenata extract. In addition, the S. crenata extract showed higher antioxidant activity than the S. hypericifolia extract. It is most likely that these effects can be explained by the higher content of individual flavonoids in the extract of S. crenata. Thus, the extract of S. crenata holds promise for more extensive research on the mechanism of its action on tumor cells.

This review addresses the use of vascular catheterization in small laboratory animals in biomedical research with an emphasis on the technological aspects of the method. The use of vascular catheters for blood sampling, drug delivery or biomonitoring improves the quality of the study (reliability and reproducibility of results) and promotes compliance with modern bioethical standards. The key factors that determine the success of the surgery and the entire study are considered with an up-to-date approach. In particular, recommendations are given on the choice of the vessel and the type and size of the catheter, depending on the characteristics of the animal and the study objectives. Catheterization of the external jugular vein of a rat is described in detail, and the fundamental stages of the procedure are the same for all major vessels of rodents. Much attention is paid to potential complications of vascular catheterization, care for catheterized animals in the postoperative period, as well as measures for maintaining the patency of the catheter and its proper functioning. The main limitations for the widespread use of catheterization in research are insufficient qualification of the surgeon and the need to use surgical equipment and microsurgical instruments.

Under conditions of steady-state hemopoiesis, nuclear factor NF-κB, in contrast to MAP kinase p38, plays an important role in the maintenance of the initial level of secretory activity of monocytes. The increase in the production of G-CSF under stress conditions (10-h immobilization) is mainly regulated by the alternative p38MARK signaling pathway via activation of p38 synthesis. It was shown that under conditions of cytostatic-induced myelosuppression, the production of protein kinase p38 in cells decreases, and it, like NF-κB, is not the main one in the production of hemopoietin by mononuclear phagocytes.

Background: Statins are currently used for secondary prevention of Coronary Heart Disease (CHD), as the lipid-lowering therapy with them is proven safe and effective. Objective: The purpose of this research is to investigate the dose-dependent effect of statins used for secondary prevention of coronary heart disease, as well as mechanisms of quantitative and qualitative changes in lipoproteins, fatty acids and cholesterol in the blood and tissues of people of both sexes. Methods: In a clinical trial (n=125, of which 89 patients belong to group 1 and 36 to group 2) and an experiment on laboratory animals (n = 100), simvastatin reduced the total level of fatty acids in blood plasma, when given in the amount that was within the therapeutic dose range. Results: This effect was achieved through a drug-induced improvement in the capacity of hepatic cells to absorb Low-density (LDL) and Very-low-density (VLDL) lipoproteins. Conclusions: Considering the formation of saturated fatty acids, statin performed better in males. With Omega-3 polyunsaturated fatty acids involved, changes in lipoproteins, cholesterol and fatty acids (liver and myocardium) were similar to those caused by small doses of a statin drug. Effects of the combination of bisoprolol and acetylsalicylic acid were completely different from those caused by the use of statin.

Suppression of the production of granulocytic CSF under the effect of 5-fluorouracyl is related to disorders in the NF-κB-, cAMP-dependent signaling pathways and MAPK cascade. These secondary messengers are involved in the regulation of functional activity of nonadherent myelokaryocytes starting from day 10 of the experiment (initial period of the hemopoietic granulocytic stem regeneration after antimetabolite challenge). Granulocytic CSF does not play essential role in the formation of colony-stimulating activity of cells of the adherent and nonadherent fractions of the bone marrow. Only cAMP-dependent pathway is involved in the regulation of the realization of the granulocytic precursor growth potential in response to the challenge.