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Abstract Background Due to non-vascularized and aneural structure, articular cartilage has limited self-repairing capacity. The aim of this study was to investigate the revitalization of inflammatory injured articular cartilage matrices by human nasal chondrocytes (hNC). Materials and methods Cartilage matrix was prepared by devitalization of articular cartilage samples obtained intraoperatively from an adult patient undergoing knee joint replacement. hNC were obtained from native tissues by enzymatic digestion with further expansion over two passages. The obtained nasal chondrocytes were used to seed decellularized scaffolds, which were then cultured in vitro for 7, 14, or 21 days in chondrogenic medium. Migration was observed by histologic staining with fast green, safranin-O, and hematoxylin and scanning electron microscopy. Biochemical analysis was performed to determine the glycosaminoglycan (GAG) and DNA content of the cartilage using dimethylmethylene blue and CyQuant Cell Proliferation Assay Kit. Results We seeded healthy and inflamed cartilage with nasal chondrocytes and found that the cells actively invade mainly pathologically altered cartilage. The results of biochemical quantitative analysis showed that the amount of DNA significantly increased by day 7 and decreased by day 14, while the quantitative values of GAGs had the opposite trend. Histological staining showed that cartilage formation occurred on day 7, intercellular spaces were filled with de novo synthesized cartilage matrix with significantly low GAG content on day 14, and newly formed GAG-rich cartilage was observed on day 21. The obtained data on cartilage regeneration were confirmed by scanning electron microscopy. Conclusions Our preliminary results showed that human nasal chondrocytes are capable of infiltrating the pathologically altered extracellular matrix of articular cartilage damaged by arthritis, thereby promoting its repair to a physiologically relevant state. Graphical Abstract

The 5-year overall survival rate for stage IV gastric cancer is lower than 10%, despite the development of systemic therapy. Conversion surgery has shown to improve survival outcomes in patients with durable clinical response on chemotherapy. We report a clinical case of a patient, who underwent conversion surgery after pembrolizumab in the third-line setting for stage IV gastric cancer. The patient did not have recurrence for 22 months after conversion surgery.

Stomach cancer is an aggressive disease and represents a global health problem. Currently, more than 50% of patients with a newly diagnosed stomach cancer have a locally advanced process that requires a combined approach – a combination of surgical treatment and neoadjuvant/perioperative therapy. In a number of patients, after neoadjuvant therapy, there is a marked or even complete regression of the tumor; the prognosis of the disease, as well as the need for adjuvant therapy in this category of patients is the subject of discussion. This paper demonstrates the clinical observation of a complete clinical, radiological and morphological response after neoadjuvant therapy (2 cycles of chemotherapy FLOT + chemoradiotherapy), as well as long-term relapse-free survival in a patient with locally advanced gastric cancer.

During the years of widespread use of radiotherapy in cancer patients, the incidence of local radiation damage to soft tissues (of varying severity) reached 30–40% or more. The experience of the radiation injury treatment department in the 70s and 80s indicates that every 2–3 beds were occupied by patients with this pathology. With the development of radiology, the introduction of new sources and techniques, the frequency of formation of such changes in the zone of irradiation fields has significantly decreased and currently stands at about 2%. The purpose of the proposed study is to demonstrate the possibilities of skin plastic surgery in this contingent of people who, as a rule, do not receive qualified care for a long time. As part of the work carried out, the time elapsed from the moment of formation of pathological changes to hospitalization in the department, on average, was 3–5 years, and was filled with ineffective conservative measures. Over the past 4 years, 60 reconstructive plastic surgeries have been performed in the radiation injury treatment department using musculoskeletal (21), skin fascial (21), skin (10) flaps, as well as plasty with local tissues (8). In the early postoperative period, the most common complication was marginal necrosis of the flap (13) which in 11 patients caused repeated operations in the form of secondary sutures or, in the case when it was not possible to perform this, plastic surgery of the defect with insular skin flaps was applied to the granulating wound. Subsequently, complete healing of the postoperative wound was noted in such patients. Thus, the proposed options for the reconstruction of radiation ulcers that developed after combined and radiation treatment of malignant neoplasms are highly effective methods that can get rid of disfiguring defects, improve the quality of life and restore social activity in almost 100% of patients.

This case report and literature review presents the imaging features of the rare pathological condition – intravenous leiomyomatosis. Correct interpretation of diagnostic imaging data and morphological verification of intravenous leiomyomatosis helps to perform timely and complete surgical treatment.

In the Russian Federation, the incidence of uterine body cancer remains the highest among malignant neoplasms of the female genital organs. Surgical treatment is the primary method for treating early stages of the disease; however, the issue of predicting metastatic lymph node involvement at the preoperative stage for performing lymphadenectomy remains debatable. The potential prognostic significance of tumor involvement in the lower uterine segment regarding lymph node metastases may justify the need to assess tumor localization before starting treatment. Magnetic resonance imaging (MRI) of the pelvic organs can help determine the involvement of the lower uterine segment in the tumor process, thereby creating a basis for expanding the scope of surgical intervention.

Space exploration is perhaps one of the most difficult tasks ever undertaken since the emergence of humankind. The International Space Station is a unique platform for advanced technology research that is not possible anywhere else. Tissue engineering in outer space, where state of the gravity can be ‘turned off’ or ‘turned on’ in the case of application of centrifuges, is a new research field with high-value goals. The microgravity conditions allow to design novel biomaterials that cannot be produced on Earth but benefit the Earth civilisation. Developing and manufacturing a biomaterial to address a space-based challenge may lead to novel biomaterials that will find important applications in medicine on Earth and/or for long-duration space missions. Today, there are only a handful of emerging biomaterials that have been tested in space, none of which have been used for their eventual function. This paper presents advances in space technology via 3D magnetic assembly: the development of synthetic bone graft constructs aboard the International Space Station during expeditions 60/61 with clear evidence of the materials' functioning in preclinical (animal) tests on Earth. The results indicate high osteoconductivity and ultimately a good rate of tissue formation by the bone grafts prepared in space.

Literature data and own experience of nutritional support in oncology has been analyzed. Nutritional support is an integral part of the management of cancer patients. Diagnosis of nutritional deficiencies is important, with an accent on early detection of muscle loss and severely malnourished patients. The main methods of nutritional support are oral nutritional supplements, enteral and parenteral nutrition, as well as dietary advice and maintenance of physical activity. Oral nutrition should always be used in the absence of absolute contraindications. The effectiveness and indications of prehabilitation measures require further study, but now it is possible to single out a category of patients in whom they can be fully implemented. It can be cases requiring neoadjuvant chemotherapy.

Aim. Computed tomographic preoperative assessment of sarcopenia in patients with gastric cancer as a part of combined treatment and the study of the relationship between sarcopenia and postoperative complications. Materials and methods. We analyzed retrospectively the data of 65 patients with morphologically verified diagnosis of stage IIA–IIIC gastric cancer. All patients at stage I of the combined treatment received neoadjuvant chemotherapy (NCT) according to the FLOT regimen, and then a gastrectomy with D2 lymph node dissection was performed. Computed tomography (CT) scans were performed before NCT initiation and 3.5 to 5 weeks after NCT completion for preoperative staging and assessment of tumor response. Using the obtained CT data, CT sarcometry was performed by calculating the cross-sectional area of skeletal muscles at the LIII vertebra level and the “skeletal-muscular index”. Postoperative complications were assessed using the Clavien–Dindo classification. Non-parametric comparison methods were used to assess the significance of differences between groups and subgroups (Wilcoxon T-test, Pearson χ2 test). Differences were considered statistically significant at p0.05. Results. We analyzed the incidence of sarcopenia in the examined patients before the NCT and immediately before the surgery. Sarcopenia was noted in 41 (63.1%) of 65 patients before NCT and in 50 (76.9%) patients after NCT, which indicates its high prevalence in patients with stomach cancer and the adverse effect of NCT on the muscular status of patients (76.9% versus 63.1%). Postoperative complications were diagnosed in 12 of 65 patients (18.5%). The rate of postoperative complications in patients with and without sarcopenia was not statistically significantly different (p=0.392); however, complications of Clavien–Dindo grade ≥IIIb occurred only in patients with sarcopenia (p0.001). Conclusion. According to our data, NCT in gastric cancer aggravates preoperative nutritional disorders. CT, as a standard method of staging and assessing the tumor response to NCT for gastric cancer, provides a tool to assess the state of muscle mass in patients before and after NCT. Preoperative sarcopenia is a risk factor for severe postoperative complications in patients with gastric cancer after gastrectomy.

There is a steady trend towards de-escalation of surgical intervention in the breast cancer treatment strategy. At first, this topic concerned the de-velopment and implementation of methods of organ-preserving operations on the mammary gland itself. With the advent of the concept and the development of the sentinel lymph node biopsy technique, the problem of performing a gentle lymphadenectomy with the prevalence of the cN0 process was solved. In recent years, the most innovative technique has begun to develop, namely targeted lymphadenectomy, which allows per- forming economical interventions on the lymph pathways with the prevalence of the cN1→ypN0 process, that is, in the presence of a verified metastatic lymph node with complete clinical resorption of metastasis during neoadjuvant polychemotherapy. The article is devoted to the de- scription of the technique of performing targeted lymphadenectomy.

Abstract Background The aim of study was to evaluate survival outcome and limb function in cancer patients with proximal limbs metastasis. Associated factors on survival outcome and limb function were identified. The comparative analysis between intramedullary nailing and prosthesis surgery in cancer patients with proximal limb metastasis was performed. Methods In this five-center retrospective study, patients diagnosed with limbs metastasis were collected. Descriptive statistics was used and log-rank test was performed to analyze the survival in subgroups. The Cox proportional hazards regression analysis was performed to identify the independent prognostic factors. The Musculoskeletal Tumor Society (MSTS) scoring system was used to evaluate limb function after surgery, and t test or analysis of variance (ANOVA) was utilized in subgroup analysis. Results A total of 316 patients with limb metastasis were included with mean age at 61.0 years. The most common primary tumor was breast, followed by renal cancer and lung cancer. The median overall survival was 24.0 months and the 1-, 3- and 5-year survival rates were 86.9%, 34.7% and 6.8%, respectively. Primary tumor type, visceral metastasis and chemotherapy were proved to be the independent prognostic factors. The mean Musculoskeletal Tumor Society (MSTS) score was 20.5, significant difference was observed in subgroup of solitary/multiple bone metastasis, with/without pathological fracture, and type of surgery. Conclusion The present study concluded that primary tumor type, visceral metastasis and chemotherapy were three factors affecting the survival of patients. Compared with intramedullary nailing, the patients underwent prosthesis surgery showed better limb function, this procedure should be encouraged in patients with indication.

In recent years, major attention has been devoted to immunotherapy in oncology. Immunotherapy is a promising treatment approach targeted to inhibit tumor growth via stimulating the body’s antitumor immune response. Purpose. The study was aimed to investigate the relationship between melanoma growth and metastasis in mice with the various schemes of Galavit treatment. Materials and methods. We used B16-F10 melanoma in F1-mouse as an experimental model. We divided animals in 3 groups: control (no therapy after tumor transplantation); immediate Galavit therapy at a dose of 5 mg/mouse after tumor transplantation; postponed Galavit therapy a week after tumor transplantation at doses of 5mg/mouse. We evaluated the rate of tumor node growth, 50-day survival rate and metastatic development after the 50th day follow-up period. Results. We revealed 80% survival rate for 50-day observation period in the control group. Interestingly, the 50-day survival rate was 60% for the animals treated with Galavit immediately after tumor inoculation. In other group with postponed Galavit therapy, survival rate was 100 %. A similar pattern was observed for tumor growth rates. Conclusion. We found that therapy with Galavit a week after tumor inoculation significantly increased the 50-day survival rate, suppressed tumor growth and the development of metastases in comparison with the control group and the group with immediate Galavit administration.

Rectal cancer (RC) is one of the leading tumor location in the structure of the incidence of malignant neoplasms in the Russian Federation and the world. And the standard approach to the treatment of patients with locally advanced forms of RC is preoperative chemo-radiotherapy (CRT) with delayed surgery. The use of such sort of approach in the recent decades has led to the reduction of the frequency of local recurrence up to 10% and even less. However, approximately a third of patients die of distant metastases. In this regard, one of the main tasks in the treatment of patients with locally advanced forms of RC with adverse prognostic factors is the prevention of distant metastasis formation. Early initiation of the systemic therapy before surgery is aimed at solving this issue. Conducting neoadjuvant chemotherapy (NCT) instead of CRT in RC treatment allows to avoid radiation reactions and injuries, occurring in some patients. Two-component oxaliplatin-containing regimens are the most well studied types of NCT in the treatment of patients with non-metastatic RC. In this connection, despite the differences in the treatment regimens and the number of cycles, a good tolerability of the method as well as no effect on the frequency of postoperative complications and in general a satisfactory results comparable to the effects of CRT were observed. The use of NCT in combination with targeted treatment modalities as well as three-component chemotherapy regimens are promising and encouraging treatment options for patients with RC with adverse prognostic factors.

Background. Bones as an organ are one of the most common targets for tumor metastasis. Currently, the number of patients undergoing surgical treatment for metastatic bone lesions is steadily increasing. In most patients, after surgical treatment, the manifestation of clinical symptoms decreases, primarily pain syndrome, which improves their quality of life. However, it should be noted that the number of patients with bone metastases who underwent revision surgery is also increasing. This article retrospectively analyzes the factors leading to revision after surgical treatment of metastases in long bones. The aim of this study was to identify factors leading to revision after surgical treatment of patients with metastases in long bones. Methods. A retrospective medical records analysis of 247 patients who underwent surgical treatment for metastases in long bones on the basis of the P.A. Herzen Moscow Research Institute of Oncology in 20062020 was performed. Of these, 33 patients underwent revision surgery. The median age was 62 years. The localization of the primary tumor was as follows: breast cancer 10 cases, kidney cancer 13, lung cancer 3, prostate cancer 2, rectal cancer 3, liver cancer and Ewings sarcoma with bone metastases 1 case each. Results. The following factors led to revision surgery: mistakes in preoperative diagnosis (3 patients); postoperative infectious complication (6 patients); dislocation of the endoprosthesis (4 patients); continued growth of solitary metastasis after osteosynthesis (5 cases); aseptic instability after intramedullary osteosynthesis (14 patients); traumatic fracture of the endoprosthesis stem (1 patient). Conclusions. Revision after surgical treatment of metastases in long bones, in addition to postoperative complications, lead to mistakes in diagnosis and incorrect choice of surgical treatment method. To reduce the risk of revision surgical interventions, a multidisciplinary approach is needed with the development of surgical treatment tactics in consultation and the use of specialized scales of oncological prognosis.

Reliable methods for identifying rodents play an important role in ensuring the success of preclinical studies. However, animal identification remains a trivial laboratory routine that is not often discussed, despite the fact that more than 6 million rodents are used in animal studies each year. Currently, there are extensive regulations in place to ensure adequate anesthesia and to reduce animal suffering during experiments. At the same time, not enough attention is paid to the comfort of rodents during routine identification procedures, which can be painful and cause some complications. In order to achieve the highest ethical standards in laboratory research, we must minimize animal discomfort during the identification phase. In this article, we discuss traumatic methods of identification and describe several painless methods for marking in long-term experimental studies. The use of non-traumatic and non-invasive methods requires the renewal of marks as they fade and additional handling of the rodents. Laboratory personnel must be trained in stress-minimizing handling techniques to make mark renewal less stressful.

Abstract Background Prostate cancer is one of the most common and socially significant cancers among men. The aim of our study was to reveal changes in miRNA expression profiles associated with lymphatic dissemination in prostate cancer and to identify the most prominent miRNAs as potential prognostic markers for future studies. Methods High-throughput miRNA sequencing was performed for 44 prostate cancer specimens taken from Russian patients, with and without lymphatic dissemination (N1 – 20 samples; N0 – 24 samples). Results We found at least 18 microRNAs with differential expression between N0 and N1 sample groups: miR-182-5p, miR-183-5p, miR-96-5p, miR-25-3p, miR-93-5p, miR-7-5p, miR-615-3p, miR-10b, miR-1248 (N1-miRs; elevated expression in N1 cohort; p

Abstract Background Carotid and vagal paragangliomas (CPGLs and VPGLs) are rare neoplasms that arise from the paraganglia located at the bifurcation of carotid arteries and vagal trunk, respectively. Both tumors can occur jointly as multiple paragangliomas accounting for approximately 10 to 20% of all head and neck paragangliomas. However, molecular and genetic mechanisms underlying the pathogenesis of multiple paragangliomas remain elusive. Case presentation We report a case of multiple paragangliomas in a patient, manifesting as bilateral CPGL and unilateral VPGL. Tumors were revealed via computed tomography and ultrasound study and were resected in two subsequent surgeries. Both CPGLs and VPGL were subjected to immunostaining for succinate dehydrogenase (SDH) subunits and exome analysis. A likely pathogenic germline variant in the SDHD gene was indicated, while likely pathogenic somatic variants differed among the tumors. Conclusions The identified germline variant in the SDHD gene seems to be a driver in the development of multiple paragangliomas. However, different spectra of somatic variants identified in each tumor indicate individual molecular mechanisms underlying their pathogenesis.

Transfer of regenerative approaches into clinical practice is limited by strict legal regulation of in vitro expanded cells and risks associated with substantial manipulations. Isolation of cells for the enrichment of bone grafts directly in the Operating Room appears to be a promising solution for the translation of biomedical technologies into clinical practice. These intraoperative approaches could be generally characterized as a joint concept of tissue engineering in situ. Our review covers techniques of intraoperative cell isolation and seeding for the creation of tissue-engineered grafts in situ, that is, directly in the Operating Room. Up-to-date, the clinical use of tissue-engineered grafts created in vitro remains a highly inaccessible option. Fortunately, intraoperative tissue engineering in situ is already available for patients who need advanced treatment modalities.

The aim: To assess the effect of COMT G1947A genetic polymorphism (val158met) on the efficacy of spinal analgesia on day 1 after laparoscopic surgery for colorectal cancer.Material and methods. In a pilot study involving 100 patients with colorectal cancer, operated through laparoscopic access, using spinal analgesia (10.0–12.5 mg of bupivacaine + 200 mcg of morphine), the frequency of COMT gene G1947A (val158met) polymorphism, the intensity of pain on day 1 after surgery, the frequency and severity of nausea, vomiting, skin itching, the need for additional analgesia have been assessed.Results. The frequency distribution of alleles val/val (25%), val/met (45%) and met/met (30%) was consisted with Hardy-Weinberg equilibrium (χ2=0.96; P>0.05) and was not significantly different from the healthy donor group. In the groups of patients with various COMT alleles of val158met polymorphism, the studied parameters also did not differ significantly.Conclusion. Study did not find significant link between spinal analgesia efficacy on day 1 after laparoscopic surgery for colorectal cancer and COMT rs4680 G1947A (val158met) polymorphism. Further research to enhance the power of the study is warranted to reach the final conclusions.

Abstract Background Carotid paragangliomas (CPGLs) are rare neuroendocrine tumors that arise from the paraganglion at the bifurcation of the carotid artery and are responsible for approximately 65% of all head and neck paragangliomas. CPGLs can occur sporadically or along with different hereditary tumor syndromes. Approximately 30 genes are known to be associated with CPGLs. However, the genetic basis behind the development of these tumors is not fully elucidated, and the molecular mechanisms underlying CPGL pathogenesis remain unclear. Methods Whole exome and transcriptome high-throughput sequencing of CPGLs was performed on an Illumina platform. Exome libraries were prepared using a Nextera Rapid Capture Exome Kit (Illumina) and were sequenced under 75 bp paired-end model. For cDNA library preparation, a TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Gold (Illumina) was used; transcriptome sequencing was carried out with 100 bp paired-end read length. Obtained data were analyzed using xseq which estimates the influence of mutations on gene expression profiles allowing to identify potential causative genes. Results We identified a total of 16 candidate genes (MYH15, CSP1, MYH3, PTGES3L, CSGALNACT2, NMD3, IFI44, GMCL1, LSP1, PPFIBP2, RBL2, MAGED1, CNIH3, STRA6, SLC6A13, and ATM) whose variants potentially influence their expression (cis-effect). The strongest cis-effect of loss-of-function variants was found in MYH15, CSP1, and MYH3, and several likely pathogenic variants in these genes associated with CPGLs were predicted. Conclusions Using the xseq probabilistic model, three novel potential causative genes, namely MYH15, CSP1, and MYH3, were identified in carotid paragangliomas.

Abstract Background CpG island methylator phenotype (CIMP) is found in 15–20% of malignant colorectal tumors and is characterized by strong CpG hypermethylation over the genome. The molecular mechanisms of this phenomenon are not still fully understood. The development of CIMP is followed by global gene expression alterations and metabolic changes. In particular, CIMP-low colon adenocarcinoma (COAD), predominantly corresponded to consensus molecular subtype 3 (CMS3, “Metabolic”) subgroup according to COAD molecular classification, is associated with elevated expression of genes participating in metabolic pathways. Methods We performed bioinformatics analysis of RNA-Seq data from The Cancer Genome Atlas (TCGA) project for CIMP-high and non-CIMP COAD samples with DESeq2, clusterProfiler, and topGO R packages. Obtained results were validated on a set of fourteen COAD samples with matched morphologically normal tissues using quantitative PCR (qPCR). Results Upregulation of multiple genes involved in glycolysis and related processes (ENO2, PFKP, HK3, PKM, ENO1, HK2, PGAM1, GAPDH, ALDOA, GPI, TPI1, and HK1) was revealed in CIMP-high tumors compared to non-CIMP ones. Most remarkably, the expression of the PKLR gene, encoding for pyruvate kinase participating in gluconeogenesis, was decreased approximately 20-fold. Up to 8-fold decrease in the expression of OGDHL gene involved in tricarboxylic acid (TCA) cycle was observed in CIMP-high tumors. Using qPCR, we confirmed the increase (4-fold) in the ENO2 expression and decrease (2-fold) in the OGDHL mRNA level on a set of COAD samples. Conclusions We demonstrated the association between CIMP-high status and the energy metabolism changes at the transcriptomic level in colorectal adenocarcinoma against the background of immune pathway activation. Differential methylation of at least nine CpG sites in OGDHL promoter region as well as decreased OGDHL mRNA level can potentially serve as an additional biomarker of the CIMP-high status in COAD.

Abstract Background Carotid body tumor (CBT) is a rare neoplasm arising from paraganglion located near the bifurcation of the carotid artery. There is great intra-tumor heterogeneity, and CBT development could be associated with both germline and somatic allelic variants. Studies on the molecular genetics of CBT are limited, and the molecular mechanisms of its pathogenesis are not fully understood. This work is focused on the estimation of mutational load (ML) in CBT. Methods Using the NextSeq 500 platform, we performed exome sequencing of tumors with matched lymph node tissues and peripheral blood obtained from six patients with CBT. To obtain reliable results in tumors with low ML, we developed and successfully applied a complex approach for the analysis of sequencing data. ML was evaluated as the number of somatic variants per megabase (Mb) of the target regions covered by the Illumina TruSeq Exome Library Prep Kit. Results The ML in CBT varied in the range of 0.09–0.28/Mb. Additionally, we identified several pathogenic/likely pathogenic somatic and germline allelic variants across six patients studied (including TP53 variants). Conclusions Using the developed approach, we estimated the ML in CBT, which is much lower than in common malignant tumors. Identified variants in known paraganglioma/pheochromocytoma-causative genes and novel genes could be associated with the pathogenesis of CBT. The obtained results expand our knowledge of the mutation process in CBT as well as the biology of tumor development.

We present an extremely rare clinical case of a 38-year-old Russian patient with multiple malignant neoplasms of the uterus and colon caused by genetically confirmed two hereditary diseases: Diamond–Blackfan anemia and Lynch syndrome. Molecular genetic research carried out by various methods (NGS, Sanger sequencing, aCGH, and MLPA) revealed a pathogenic nonsense variant in the MSH6 gene: NM_000179.2: c.742C>T, p.(Arg248Ter), as well as a new deletion of the chromosome 15’s locus with the capture of 82,662,932–84,816,747 bp interval, including the complete sequence of the RPS17 gene. The lack of expediency of studying microsatellite instability in endometrial tumors using standard mononucleotide markers NR21, NR24, NR27, BAT25, BAT26 was demonstrated. The estimated prevalence of patients with combination of Diamond–Blackfan anemia and Lynch syndrome in the world is one per 480 million people.

Photodynamic therapy (PDT) is an effective treatment for a number of solid malignancies. In this work, the antitumor efficacy of photodynamic therapy for murine B16 melanoma with intravenous administration of a new photosensitizer (PS) based on the chlorin e6 conjugate with a prostate-specific membrane antigen (PSMA) was studied in vivo. We have previously published the data obtained in the first part of the study: the dynamics of PS accumulation in the tumor and surrounding tissues and the antitumor efficacy of the photodynamic therapy, which was evaluated by the regression parameters and morphological characteristics of the tumors—including by the complete regression of the tumors, the absolute growth rate of the tumors among the mice with continued tumor growth, and an increase in life expectancy compared to the control. The criterion for a complete cure was the absence of signs of tumor recurrence within 90 days after therapy. The conducted studies demonstrated the high efficiency of the new photosensitizer for the photodynamic therapy of B16 melanoma. This article presents a continuation of this work, including histological studies of the zones exposed to laser irradiation on the 21st day after treatment and an assessment of the therapeutic potential of photodynamic therapy for the destruction of tumor cells. Pathological studies in the zones of photodynamic exposure revealed that the effectiveness of the PDT depended on the PS dose and the laser irradiation parameters.

Prostate cancer (PC) is one of the most common cancers among men worldwide, and advanced PCs, such as locally advanced PC (LAPC) and castration-resistant PC (CRPC), present the greatest challenges in clinical management. Current indicators have limited capacity to predict the disease course; therefore, better prognostic markers are greatly needed. In this study, we performed a bioinformatic analysis of The Cancer Genome Atlas (TCGA) datasets, including RNA-Seq data from the prostate adenocarcinoma (PRAD; n = 55) and West Coast Dream Team – metastatic CRPC (WCDT-MCRPC; n = 84) projects, to evaluate the transcriptome changes associated with progression-free survival (PFS) for LAPC and CRPC, respectively. We identified the genes whose expression was positively/negatively correlated with PFS. In LAPC, the genes with the most significant negative correlations were ZC2HC1A, SQLE, and KIF11, and the genes with the most significant positive correlations were SOD3, LRRC26, MIR22HG, MEG3, and MIR29B2CHG. In CRPC, the most significant positive correlations were found for BET1, CTAGE5, IFNGR1, and GIMAP6, and the most significant negative correlations were found for CLPB, PRPF19, ZNF610, MPST, and LINC02001. In addition, we performed a gene network interaction analysis using STRINGdb, which revealed a significant relationship between genes predominantly involved in the cell cycle and characterized by upregulated expression in early recurrence. Based on the results, we propose several genes that can be used as potential prognostic markers.

Head and neck paragangliomas (HNPGLs) are rare neuroendocrine tumors that have a high degree of heritability and are predominantly associated with mutations in ten genes, such as SDHx, SDHAF2, VHL, RET, NF1, TMEM127, MAX, FH, MEN2, and SLC25A11. Elucidating the mutation prevalence is crucial for the development of genetic testing. In this study, we identified pathogenic/likely pathogenic variants in the main susceptibility genes in 102 Russian patients with HNPGLs (82 carotid and 23 vagal paragangliomas) using whole exome sequencing. Pathogenic/likely pathogenic variants were detected in 43% (44/102) of patients. We identified the following variant distribution of the tested genes: SDHA (1%), SDHB (10%), SDHC (5%), SDHD (24.5%), and RET (5%). SDHD variants were observed in the majority of the patients with bilateral/multiple paragangliomas. Thus, among Russian patients with HNPGLs the most frequently mutated gene was SDHD followed by SDHB, SDHC, RET, and SDHA.

Abstract Background Carotid body tumor (CBT) is a form of head and neck paragangliomas (HNPGLs) arising at the bifurcation of carotid arteries. Paragangliomas are commonly associated with germline and somatic mutations involving at least one of more than thirty causative genes. However, the specific functionality of a number of these genes involved in the formation of paragangliomas has not yet been fully investigated. Methods Exome library preparation was carried out using Nextera® Rapid Capture Exome Kit (Illumina, USA). Sequencing was performed on NextSeq 500 System (Illumina). Results Exome analysis of 52 CBTs revealed potential driver mutations (PDMs) in 21 genes: ARNT, BAP1, BRAF, BRCA1, BRCA2, CDKN2A, CSDE1, FGFR3, IDH1, KIF1B, KMT2D, MEN1, RET, SDHA, SDHB, SDHC, SDHD, SETD2, TP53BP1, TP53BP2, and TP53I13. In many samples, more than one PDM was identified. There are also 41% of samples in which we did not identify any PDM; in these cases, the formation of CBT was probably caused by the cumulative effect of several not highly pathogenic mutations. Estimation of average mutation load demonstrated 6–8 mutations per megabase (Mb). Genes with the highest mutation rate were identified. Conclusions Exome analysis of 52 CBTs for the first time revealed the average mutation load for these tumors and also identified potential driver mutations as well as their frequencies and co-occurrence with the other PDMs.

Abstract Background Colorectal cancer (CRC) is a common cancer worldwide. The main cause of death in CRC includes tumor progression and metastasis. At molecular level, these processes may be triggered by epithelial-mesenchymal transition (EMT) and necessitates specific alterations in cell metabolism. Although several EMT-related metabolic changes have been described in CRC, the mechanism is still poorly understood. Results Using CrossHub software, we analyzed RNA-Seq expression profile data of CRC derived from The Cancer Genome Atlas (TCGA) project. Correlation analysis between the change in the expression of genes involved in glycolysis and EMT was performed. We obtained the set of genes with significant correlation coefficients, which included 21 EMT-related genes and a single glycolytic gene, HK3. The mRNA level of these genes was measured in 78 paired colorectal cancer samples by quantitative polymerase chain reaction (qPCR). Upregulation of HK3 and deregulation of 11 genes (COL1A1, TWIST1, NFATC1, GLIPR2, SFPR1, FLNA, GREM1, SFRP2, ZEB2, SPP1, and RARRES1) involved in EMT were found. The results of correlation study showed that the expression of HK3 demonstrated a strong correlation with 7 of the 21 examined genes (ZEB2, GREM1, TGFB3, TGFB1, SNAI2, TWIST1, and COL1A1) in CRC. Conclusions Upregulation of HK3 is associated with EMT in CRC and may be a crucial metabolic adaptation for rapid proliferation, survival, and metastases of CRC cells.

Abstract Background Neuropilin and tolloid-like 2 (NETO2) is a single-pass transmembrane protein that has been shown primarily implicated in neuron-specific processes. Upregulation of NETO2 gene was also detected in several cancer types. In colorectal cancer (CRC), it was associated with tumor progression, invasion, and metastasis, and seems to be involved in epithelial-mesenchymal transition (EMT). However, the mechanism of NETO2 action is still poorly understood. Results We have revealed significant increase in the expression of NETO2 gene and deregulation of eight EMT-related genes in CRC. Four of them were upregulated (TWIST1, SNAIL1, LEF1, and FOXA2); the mRNA levels of other genes (FOXA1, BMP2, BMP5, and SMAD7) were decreased. Expression of NETO2 gene was weakly correlated with that of genes involved in the EMT process. Conclusions We found considerable NETO2 upregulation, but no significant correlation between the expression of NETO2 and EMT-related genes in CRC. Thus, NETO2 may be involved in CRC progression, but is not directly associated with EMT.

РезюмеМетоды иммунокоррекции, основанные на введении в организм аутологичных лейкоцитов, активированных в условиях in vitro различными иммуномодуляторами, получают все более широкое распространение. Ранее проведенные нами исследования позволяют утверждать об эффективности применения иммунотерапии лимфоцитами, активированными in vitro с помощью цитокинов для лечения онкологических больных и хорошей переносимости данной терапии. В данной статье рассмотрены варианты применения препаратов рекомбинантного интерферона гамма (IFN-γ) и рекомбинантного фактора некроза опухоли-альфа-тимозина-альфа1 (TNF-α-Т) для получения цитокин-индуцированных киллеров (ЦИК) человека in vitro Целью исследования являлась оценка возможности применения препаратов IFN-γ и TNF-α-Т в протоколах получения ЦИК человека в условиях in vitro. Материалы и методы. Мононуклеары (МНК), выделенные из крови 15 доноров, культивировали в условиях СО2-инкубатора до 12 суток. Для оценки токсичности IFN-γ и TNF-α-Т в отношении лимфоцитов in vitro концентрации препаратов были выверены экспериментальным путем с учетом контроля уровня жизнеспособности клеток. Уровни цитокинов в супернатантах на этапах культивирования клеток определяли методом иммуноферментного анализ (ИФА). До и после культивирования оценивали наличие адгезии и гранулярности изучаемых лимфоцитов, фенотип клеток изучали методом проточной цитометрии, оценку уровня жизнеспособности активированных лимфоцитов проводили в камере Горяева. Результаты. Проведен сравнительный анализ способов культивирования лимфоцитов в различных средах, содержащих: только IFN-γ (группа 1); IFN-γ, интерлейкин-2 (IL-2) и интерлейкин-15 (IL-15) (группа 2); только TNF-α-Т (группа 3); TNF-α-Т, IL-2 и IL-15 (группа 4) и только IL-2 и IL-15 (группа контроля). При активации МНК в среде, содержажей IFN-γ, IL-2 и IL-15 или в среде, содержащей TNF-α-Т, IL-2 и IL-15, отмечена общая тенденцию к значительному увеличению показателей Т-цитотоксических клеток (CD3+CD8+), NKТ-клеток (CD3+CD16+CD56+), активированных лимфоцитов (HLA-DR+), активированных Т-лимфоцитов (CD3+HLA-DR+), и маркеров активации на всех лимфоцитах (CD38+) и на Т-клетках (CD3+CD38+). Заключение. Оценена возможность применения препаратов IFN-γ и TNF-α-Т для получения ЦИК человека in vitro. Полученные данным способом активированные цитотоксические лимфоциты могут применяться для фундаментальных исследований по выявлению новых закономерностей и морфо-функциональных особенностей активированных лимфоцитов человека, а также для проведения адоптивной иммунотерапии онкологическим больным.AbstractMethods of immunocorrection based on the introduction into the body of autologous leukocytes activated in vitro by various immunomodulators are becoming increasingly widespread. Our previous studies suggest that the use of immunotherapy with lymphocytes activated in vitro using cytokines is effective for the treatment of cancer patients and that this therapy is well tolerated. This article discusses the use of drugs: recombinant interferon-gamma (IFN-γ) and recombinant tumor necrosis factor-alpha-thymosin-alpha1 (TNF-α-T) for the production of human cytokine-induced killers (CIK) in vitro. The purpose of the study was to assess the possibility the use of IFN-γ and TNF-α-T drugs in protocols for obtaining human CIK in vitro. Materials and methods. Isolated mononuclear cells from 15 donors were cultured in a CO2 incubator for 12 days and the morpho-functional characteristics of lymphocytes were studied. To assess the toxicity of IFN-γ and TNF-α-T towards lymphocytes in vitro, the concentrations of the drugs were adjusted experimentally, taking into account the control of the level of cell viability. The levels of cytokines in the supernatants at the stages of cell culture were determined by enzyme-linked immunosorbent assay (ELISA). Before and after cultivation, the presence of adhesion and granularity of the studied lymphocytes was assessed, the phenotype of the cells was studied by flow cytometry, and the level of viability of activated lymphocytes was assessed in a Goryaev chamber. Results. A comparative analysis of methods for cultivating lymphocytes in different cultures was carried out. We studied media containing: only IFN-γ (group 1); IFN-γ, interleukin-2 (IL-2) and interleukin-15 (IL-15) (group 2); TNF-α-T only (group 3); TNF-α-T, IL-2 and IL-15 (group 4) and only IL-2 and IL-15 (control group). When mononuclear cells are activated in a medium containing IFN-γ, IL-2 and IL-15 or in a medium containing TNF-α-T, IL-2 and IL-15, there is a general tendency towards a significant increase in T-cytotoxic cells (CD3 +CD8+), NKT cells (CD3+CD16+CD56+), activated lymphocytes (HLA-DR+), activated T lymphocytes (CD3+HLA-DR+), and activation markers on all lymphocytes (CD38+) and on T cells ( CD3+CD38+). Conclusion. The possibility of using IFN-γ and TNF-α-T to obtain human CIK in vitro was assessed. Activated cytotoxic lymphocytes obtained by this method can be used for fundamental research to identify new patterns and morpho-functional characteristics of activated human lymphocytes, as well as for adoptive immunotherapy for cancer patients.

Older age is one of the main risk factors for cancer development. The incidence of prostate cancer, as a multifactorial disease, also depends upon demographic factors, race, and genetic predisposition. Prostate cancer most frequently occurs in men over 60 years of age, indicating a clear association between older age and disease onset. Carcinogenesis is followed by the deregulation of many genes, and some of these changes could serve as biomarkers for diagnosis, prognosis, prediction of drug therapy efficacy, as well as possible therapeutic targets. We have performed a bioinformatic analysis of a The Cancer Genome Atlas (TCGA) data and RNA-Seq profiling of a Russian patient cohort to reveal prognostic markers of locally advanced lymph node-negative prostate cancer (lymph node-negative LAPC). We also aimed to identify markers of the most common molecular subtype of prostate cancer carrying a fusion transcript TMPRSS2-ERG. We have found several genes that were differently expressed between the favorable and unfavorable prognosis groups and involved in the enriched KEGG pathways based on the TCGA (B4GALNT4, PTK6, and CHAT) and Russian patient cohort data (AKR1C1 and AKR1C3). Additionally, we revealed such genes for the TMPRSS2-ERG prostate cancer molecular subtype (B4GALNT4, ASRGL1, MYBPC1, RGS11, SLC6A14, GALNT13, and ST6GALNAC1). Obtained results contribute to a better understanding of the molecular mechanisms behind prostate cancer progression and could be used for further development of the LAPC prognosis marker panel.

Improvement of colon cancer diagnosis is a very important medical problem. Methods of fluorescence diagnosis (FD) with 5-aminolevulinic acid (5-ALA) have been used in detection of early cancer on the surface of mucosa. The objective of the study was to estimate the effectiveness of FD with 5-ALA in the detection of malignant lesions in the colon. From 2008 to 2010 full examinations have been performed in 78 patients suffering from colon polyps. For the study we used drug Alasens based on 5-ALA. Fluorescence colonoscopy with Alasens was made using the visual assessment of fluorescence images in combination with local fluorescence spectroscopy. The results of FD were compared with those of final histopathological study. The application of the method during colonoscopy allows specifying diagnosis, identifying areas of colon tumors transformation, and also showing a place for targeted biopsy. Sensitivity and specificity of FD with Alasens were 94.9% and 62.5%, respectively. The application of local fluorescence spectroscopy as a step of the combined approach allows increasing the specificity of fluorescence colonoscopy from 62.5% up to 93.7%.