2025, volume 21

The article is dedicated to Professor Valentin S. Speransky (1925–2015), Doctor of Medical Sciences, Honored Scientist of the Russian Federation, who headed the Department of Human Anatomy from 1967 to 1996. The paper describes the main stages of the researcher’s scientific biography, who devoted more than 40 years of his life to this department. Outstanding anatomical scientist of our time, Professor Valentin S. Speransky made a significant contribution to the development of both the department and anatomy in general. The craniological school established by him is one of the leading ones in Russia and has gained recognition abroad.

The article is dedicated to the 75th anniversary of the birth of an outstanding representative of Saratov surgical school in Russia, a talented scientist and teacher, Head of General Surgery Department at V.I. Razumovsky Saratov State Medical University, Doctor of Medical Sciences, Professor Yuri G. Shapkin.

Objective: to create and evaluate the potential of an in vitro bioengineered model of the bone marrow endothelial niche for the support and activation of human hematopoietic stem cells. Material and methods. Human lymphocytes isolated from peripheral blood mononuclear cells were used. Experimental batch of platforms with various activated surfaces on the wells of 24-well plates were fabricated, namely: fibronectin only, poly-D-lysine only, fibronectin and poly-D-lysine, poly-D-lysine and anti-CD3 antibodies. During the 7-days cultivation, the cell populations state was inspected by direct microscopy, the viability was assessed via trypan blue staining, the adhesive activity assay was performed. Results. Under experimental conditions, lymphocyte viability was recorded at a high level of >80% for all test platforms and in control, respectively, with maintained typical for lymphocytes morphology. The use of the 24T_FN platform with the fibronectin activator showed non-uniform distributed cell islands and the formation of large conglomerates. The 24T_pDl platform with poly-D-lysine coating demonstrated a high degree of 2D uniformity with the formation of separate clusters within 50–100 cells. The adhesive activity on day 7 of the experiment for the platform 24T_FN and platform 24T_pDl was in the range of 60–70%, three times higher than the analogous data for control samples. The combined 24T_pDl@AbCD3 platform coated with poly-D-lysine in combination with anti-CD3 antibodies demonstrated the highest rates of cell survival, adhesion activity, and the 2-D uniformity. Conclusion. Herein, we demonstrated the capability of long-term monolayer subcultivation of human lymphocytes with high adhesive activity on the in vitro model synthetic platform based on the poly-D-lysine surface activator in combination with antibodies with affinity to CD3+ T cells.

Objective: adaptation of genome reprogramming technology based on laser transfection to create biomedical cell products. Material and methods. We developed optotransfection platforms based on multi-well culture plates with the working surface coated with gold nanoparticle monolayers. Model cell experiments on the delivery of the molecular agent propidium iodide were conducted using the mice macrophage cells, line RAW 264.7. Transfection of cell monolayers adhered to 24-well plate platforms with gold nanostar layers was carried out via single 2-D scanning with a pulsed laser and narrowly focused beam. The viability and efficiency of cell modification were assessed by direct microscopy and metabolic tests. Results. The optimal operating modes for laser optotransfection of RAW 264.7 mouse macrophage cells were determined, namely: pulse energy 1.4 μJ, pulse duration 200 ns, pulse frequency 10 kHz, scanning speed 0.025 m/s. The minimum starting monolayer confluency was 40%, the transfection efficiency was 78±7.6%, and the cell viability was 84±9.3%. Conclusion. Under controlled model in vitro experiments, it has been demonstrated that fine adjustment of optotransfection parameters enables reproducible results of genetic modification in lymphocytelike cells, achieving high levels of both efficiency and viability.

Objective: identify the highly relevent molecular cytogenetic technologies for the analysis of chromosomal disorders aimed at robust diagnosis and monitoring of socially significant human diseases. Review methodology. The systematic review was performed on the PRISMA (Preferred reporting items for systematic reviews and meta-analyses) methodology applying the PubMed, eLibrary, Google Scholar, CyberLeninka databases. The search period was from 2000 to 2025. 50 sources were used to write the review. Conclusion. A review of scientific publications suggests a significant potential of the development of molecular genetic technologies and optical imaging methods to improve the quality of diagnostic laboratory assays. Analysis of studies focused on the chromosomal disorders determination revealed the emergence of new technologies (chromosomal microarray analysis, optical genome mapping) in the diagnosis of previously poorly studied hereditary diseases. Recent studies in the field of microscopic technology related to the development of high-resolution methods for direct luminescent visualization of molecular cytogenetic data, are aimed at the fundamental improve of the identification of predictors for numerous human chromosomal diseases. Modern developmental trends in the bioengineering and nanobiotechnology are aimed at designing specialized multicolor molecular probes and hybrid nano-sized labels, which increase time- and cost efficacy of chromosomal analysis, thus making it convinient for large-scale screening studies.

Objective: to develop a simple and robust technology for DNA targets detection for the diagnosis of socially significant diseases, based on a colorimetric aggregation test using gold nanoparticles. Material and methods. Gold nanospheres were synthesized using liquid-phase chemistry methods. The physicochemical parameters of the nanoparticles were controlled by absorption spectroscopy, dynamic light scattering, and electron microscopy. The rational design of the specific sequences of target genomic fragments and oligonucleotides included in the test system, was carried out taking into account the thermodynamic constants and using genomic databases. The analytical validation of the test system was carried out using a model for detecting the DNA of Babesia species, the causative agent of human and animal pyroplasmosis, by cross-comparing the characteristics with a polymerase chain reaction with electrophoretic results detection and in a "real-time" format. Results. A pilot test system has been developed for the specific detection of up to 10 ng of DNA in blood samples by visual discrimination of the color of the reaction mixture, within analysis duration about 30 min. The size effect of nanoparticles in the range from 15 to 60 nm on the minimum detection limit of DNA targets has been revealed. Conclusion. This paper presents the results of the development and analytical validation of a pilot version of an express test system for DNA diagnostics based on gold nanoparticle tags and visual color discrimination of the reaction mixture.

Objective: to develop and validate a chromatograph mass spectrometric method for the quantitative determination of cucurbitacin B in mouse plasma and liver, kidney, and heart homogenates, and to evaluate the blood cucurbitacin B levels and concentration dynamics following a single oral administration. Material and methods. An Agilent Technologies 1260 Infinity II high-performance liquid chromatograph and an AB Sciex QTrap 3200 MD mass spectrometer were used to develop the method for the quantitative determination of cucurbitacin B. Chromatographic separation was performed on an Agilent InfinityLab Poroshell 120 EC-C18 analytical column with a gradient mobile phase. Mouse plasma sample preparation consisted of protein precipitation with methanol. Results. To study the pharmacokinetics of the components of the Grapevine officinalis extract, a bioanalytical method for determining cucurbitacin B in mouse plasma using high performance liquid chromatography with tandem mass spectrometry was developed, and cucurbitacin B identification parameters were selected (lower limit of quantification). Conclusion. To determine the pharmacokinetic parameters of cucurbitacin B, it is necessary to search for other analytes – cucurbitacin B metabolites. Consequently, it is impossible to construct a pharmacokinetic curve for cucurbitacin B. The metrological characteristics of the method allow it to be used for the analytical portion of pharmacokinetic studies of both individual substances and plant extracts.

Objective: to evaluate the effect of local application of dental gel with microencapsulated vitamin D on gingival microcirculation parameters in experimental periodontitis in white rats. Material and methods. A total of 60 white rats were studied (15 in each of the 4 groups): control group – intact animals; comparison group – rats with periodontitis at week 3; placebo group – rats with periodontitis treated with a gel containing microcapsules of silver nanoparticles 0.1; experimental group – rats with periodontitis that had a gel applied to their gum surface containing microcapsules with vitamin D. Microcirculation was investigated using laser Doppler flowmetry on day 21 of the experiment. Results. At the week 3 of the experiment, the comparison group of animals showed an increase in perfusion and a change in both active and passive mechanisms of blood flow modulation. The gel containing AgNP partially reduced the perfusion index by 13% and moderately corrected endothelial-dependent fluctuations, compared to the animals with periodontitis. The use of a gel containing vitamin D in microcapsules as an active component in the experimental group of animals was accompanied by a 15% decrease in the perfusion index compared to the animals with periodontitis at 3 weeks. The amplitude of endothelial oscillations in the experimental group decreased by 42% compared to the comparison group. Neurogenic and myogenic fluctuations in the group treated with a vitamin D-containing gel were 47 and 43% lower, respectively, compared to the periodontitis-affected animals at 3 weeks. Conclusion. The use of a gel containing vitamin D in microcapsules normalizes the microcirculatory parameters of the gums in rats with experimental periodontitis.

Objective: to identify changes in the regulatory T cell (Treg) pool and the expression level of inducible T-cell costimulator (ICOS) and programmed cell death protein (PD-1) on their membrane in metastatic lymph nodes in colorectal cancer (CC). Material and methods. The main group consisted of 105 patients with stage III CC. The clinical comparison group consisted of non-neoplastic pathology (developmental anomalies, stomas, diverticular disease of the colon). The Treg pool and the expression of PD-1 and ICOS were studied using flow cytometry. Results. In metastatic lymph nodes with RTC, the content of Treg increases by 3.5 times (p<0.001), the relative number of naive Treg decreases (p<0.001), the number of central memory Treg expressing CD197 on their membrane increases almost 2 times (p<0.001), and effector memory cells with the CD45RA–CD197– phenotype increase 3.6 times (p<0.001). The number of PD-1−ICOS+ Treg in the total population of CD3+ lymphocytes increases by 3.1 times (p<0.001). The number of Treg with the CD19−CD3+CD4+CD25highCD127−PD-1+ICOS+ phenotype increases by 1.5 times (p<0.001) compared to the control. Conclusion. In patients with RTC, the number of naive Treg in metastatically affected lymph nodes decreases, the relative content of central and effector memory Treg increases, the number of Treg expressing ICOS increases, as well as those simultaneously expressing both the inhibitory protein PD-1 and the stimulating molecule ICOS.

Objective: to identify the most accurate craniometric scheme for determining the anatomical location of the lateral sulcus of the brain in adult men with different skull shapes. Material and methods. The study included computed tomograms of 257 adult men, of whom 113 belonged to the I period of adulthood (21–35 years), and 144 to the II period of adulthood (36–60 years). Methods of craniocephalometric analysis were used to assess the localization of the lateral sulcus. Craniometric schemes were constructed according to the techniques of Taylor – Haughton, Broca, Krönlein (Kocher – Keen), and Rhoton. Results. The most common skull shape among the study participants was brachycranial (46,9% among men of the I period of adulthood and 56,2% among men of the II period of adulthood). In dolichocrania, the location of the lateral sulcus best corresponded to the Rhoton method (82%; p=0.04 – men of the I period of adulthood, p=0.015 – men of the II period of adulthood). In men with a brachycranial skull shape, the greatest correspondence was demonstrated by the Taylor-Haughton method (78%, p=0.020 – men of the I period of adulthood, p=0.003 – men of the II period of adulthood), and with a mesocranial shape – the Krönlein method (p>0.05). Conclusion. Optimal schemes were identified for determining the location of the lateral sulcus of the brain using computed tomography in men with dolichocranial and brachycranial head shapes. For men with mesocrania, an optimal scheme was not revealed, which makes it necessary to further study acceptable approaches for this craniological group.