[Predictors of therapeutic efficacy of immune response checkpoint inhibitors in clear cell renal cell carcinoma].

Immunotherapy in oncologic diseases involves the use of drugs which stimulate the immune system and indirectly suppress tumor cells growth. These agents have expanded the treatment options for cancer patients. Despite the impressive success achieved in the development of immune checkpoint inhibitors (ICIs) and subsequent approval in a broader spectrum of malignant tumors, most patients are not responded the therapy. Currently available predictive markers of efficacy are nonspecific. However, microRNAs are of particular interest, which regulate gene expression and are involved in the carcinogenesis and therapy resistance. Therefore, it is clear that for the most efficient and cost-effective use of ICIs, it is important to have validated biomarkers that will accurately predict the therapeutic response. The published results on molecular genetic changes in patients with renal cell carcinoma (RCC) were analyzed and summarized in order to determine possible prognostic biomarkers when prescribing ICI therapy.

Transtraumatic Epidural Electrostimulation of the Spinal Cord in a Pig Model.

The effect of transtraumatic epidural electrostimulation (TEES) above (T5) and below (L2) spinal cord injury in the lower thoracic region (T8-T9) in combination with treadmill exercise in pigs was evaluated using electrophysiological examination methods and behavioral tests. Two weeks after spinal cord injury, motor evoked potentials of m. soleus were recorded during electrostimulation at the level of T5 and L2 segments, which indicated activation of spinal cord structures above and below the focus of injury. After 6 weeks of TEES in combination with physical training, restoration of the characteristics of M-response and H-reflex of the soleus muscle in response to stimulation of the sciatic nerve, improvement of joint mobility, and appearance of voluntary motor activity in the hindlimbs were observed. Neuromodulation with TEES had been proven to be an effective way to stimulate posttraumatic spinal cord regeneration and can be used in the development of a neurorehabilitation protocol for patients with spinal cord injury.

Exosomal miRNA-146a is downregulated in clear cell renal cell carcinoma patients with severe immune-related adverse events.

Here we report the results of the pilot project of exosomal miRNA expression levels in clear cell renal cell carcinoma (ccRCC) patients with different clinical response to ICIs (nivolumab) and treatment related toxicity. Immune-related adverse events (irAEs) are a major cause of immune checkpoint inhibitors cancellation and therapy failure. Modern studies demonstrate evidence that exosomes are of great importance in the formation of tumor resistance to ICIs drugs and therapy. We performed exosomal miRNA-146a expression analysis using qPCR on 86 ccRCC patients and revealed a statistically significant (p = 0.01) decreased expression level in ccRCC patients with CTCAE grade 3-4 (M±SEM 1.71 ± 0.13) compared to CTCAE grade 0-2 group (M±SEM 2.30 ± 0.24). The expression levels of miRNA-126, miRNA-218 and miRNA-410 did not show statistically significant differences in the comparison groups (p > 0.05). Association analysis of rs2910164 in the miRNA-146a gene demonstrated that CC genotype and C allele carriers had higher risk of developing severe irAEs (p = 0.03, OR = 6.12; p = 0.01, OR = 2.42, respectively) compare with GG and GC carriers. That is the first attempt to identify biomarkers of ICIs treatment efficacy for ccRCC in the Volga-Ural region based on exosomal miRNAs analysis.

Methylation and expression levels of microRNA-23b/-24-1/-27b, microRNA-30c-1/-30e, microRNA-301a and let-7g are dysregulated in clear cell renal cell carcinoma.

BACKGROUND: Renal cell carcinoma is the most common form of kidney cancer in adults. DNA methylation of regulatory sequences at the genomic level and interaction between microRNAs and the messenger RNAs of target genes at the posttranscriptional level contribute to the dynamic regulation of gene activity. Aberrations in these mechanisms can result in impaired functioning of cell signaling pathways, such as that observed in malignant tumors. We hypothesized that microRNA genes methylation may be associated with renal cancer in patients. METHODS AND RESULTS: We examined methylation levels of 22 microRNA genes in tumor and normal kidney tissue of 30 patients with TNM Stage III clear cell renal cell carcinoma using a pathway-specific real-time polymerase chain reaction array (EpiTect Methyl II PCR Arrays, Qiagen). MicroRNA expression analysis by quantitative polymerase chain reaction was also performed. Significant differences in methylation levels were found in two genes and in two clusters of microRNA genes. MicroRNA-23b/-24-1/-27b, microRNA -30c-1/-30e and let-7 g was hypermetylated in clear cell renal cell carcinoma tissue, microRNA -301a was hypomethylated in tumor compared with the adjacent normal tissues. Expression of microRNA-301a, microRNA-23b in the clear cell renal cell carcinoma tissues was significantly overexpressed when compared with the adjacent normal tissues and let-7 g was significantly downregulated in tumor. CONCLUSIONS: Our results may indicate the contribution of microRNA-301a, microRNA-23b and let-7 g in the pathogenesis of renal cancer, but further studies are needed to determine the functional significance of the detected changes.

[The potential role of mirnas in the pathogenesis of hemorrhagic fever with renal syndrome].

Hemorrhagic fever with renal syndrome (HFRS) is an acute natural focal viral disease caused by viruses of the genus hantavirus, characterized by damage to small blood vessels, kidneys, lungs and other organs of a person. MicroRNAs (miRNAs) are 18-22 nucleotide endogenously expressed RNA molecules that inhibit gene expression at the post-transcriptional level by binding to the 3-untranslated region of the target mRNA. It has been proven that miRNAs play a significant role in various biological processes, including the cell cycle, apoptosis, cell proliferation and differentiation. It has been proven that miRNAs may be involved in the pathogenesis of infectious diseases, including HFRS. Hantavirus infection predominantly affects endothelial cells and causes dysfunction of the endothelium of capillaries and small vessels. It is known that the immune response induced by Hantavirus infection plays an important role in disrupting the endothelial barrier. In a few studies, both in vitro and in vivo, it has been shown that endothelial dysfunction and the immune response after infection with Hantavirus can be partially regulated by miRNAs by acting on certain genes. Most of the miRNAs is expressed within the cells themselves. However, in some biological fluids of the human body, for example, plasma or blood serum, numerous miRNAs, called circulating miRNAs, have been found. Circulating miRNAs can be secreted by cells into human biological fluids as part of extracellular vesicles as exosomes or be part of an RNA-bound protein complex as miRNA-Argonaute 2 (Ago2). These miRNAs are resistant to nucleases, which makes them attractive as potential biomarkers in various human diseases. There is no specific antiviral therapy for HFRS, and the determination of laboratory parameters that are used to diagnose, assess the severity, and predict the course of the disease remains a challenge due to the peculiarities of the pathophysiology and clinical course of the disease. Studying the role of miRNAs in HFRS seems to be expedient for the development of specific and effective therapy, as well as for use as diagnostic and prognostic biomarkers (in relation to circulating miRNAs).

[Analysis of the results of ERAS protocol in real-life clinical practice AFTER radical cystectomy (the first prospective multicenter study in Russia)].

AIM: to carry out a multicenter prospective analysis of the results of the ERAS protocol in patients undergoing radical cystectomy in real-life clinical practice. The aims of the study were to assess the complication and mortality rate after radical cystectomy using the ERAS protocol and to assess how often ERAS protocol was imple- mented. MATERIALS AND METHODS: a multicenter study was carried out in 4 clinics in Russia. A total of 134 patients who underwent radical cystectomy in 2017 were prospectively analyzed. Open and laparoscopic radical cystectomy was performed in 35 (26.1%) and 99 (73.9%) patients, respectively. Bricker procedure prevailed as a method for urine derivation (91.7%). Complication and mortality rate, and each principle of ERAS protocol was analyzed both in the general sample of patients and separately for open and laparoscopic radical cystectomy. RESULTS: length of hospitalization before the radical cystectomy was 1 (1-2) day. The median duration of surgery was 260 (205-300) minutes, median blood loss was 300 (200-400) ml. The median of the patients time in ICU was 1 (0-2) day. A total of 95 (70%) complications were recorded in the 90-day period after the surgery, including Clavien I-II category in 52 (38.8%) cases and Clavien III-IV in 43 (32%) cases. Of these, gastrointestinal tract complications were predominated. Gastroparesis requiring a nasogastric tube was observed in 16 (11.9%) patients. Ileus developed in 43 (32.1%) cases, and 22 patients (16.4 %) were managed conservatively; however, 21 patients (15.7%) undergone to reoperation. A 90-days mortality reached 5.2% and the main causes included multiple organ failure as a complication of peritonitis, acute heart failure after myocardial infarction and massive bleeding. Re-hospitalization rate was 9.7% (n=13). Length of stay was 12 (9-16) days. According to univariate and multivariate analysis, an absence of antibacterial prophylaxis, a history of coronary heart disease and the patients age more than 75 years were predictors of an increased complication rate. A 30-days mortality rate is 5.2%, and re-hospitalization was required in 9.7% (n=13) cases. An average length of stay was 12 (9-16) days. Frequency of implementation of ERAS protocol in each of the participating clinic varied. Open and laparoscopic radical cystectomy have insignificant differences in some intra- and postoperative parameters, but, in general, both approaches are comparable in terms of complications, mortality, and length of stay. CONCLUSION: 1. Despite the use of the ERAS protocol, radical cystectomy has a high frequency of complications (up to 70%); most of them are Clavien I-II. A 30-days mortality rate is 5.2%, and re-hospitalization is required in 9.7% cases. 2. Univariate and multivariate analysis showed that an absence of antibacterial prophylaxis, a coronary heart disease and the patients age more than 75 years are predictors of an increased complication rate. 3. Open and laparoscopic radical cystectomy have insignificant differences in some intra- and postoperative parameters, but, in general, both approaches are comparable in terms of complications, mortality, and length of stay. 4. To obtain more convincing data on the ERAS protocol after radical cystectomy, long-term studies are required.

[A weak association of 677 C>T polymorphism in MTHFR with recurrent embryonic loss].

Early (embryonic) pregnancy loss before 10 week of gestation (wg) could also be related with endometrial receptivity as well as with gene expression regulation in developed embryo. Methylation of genome is a key process in the gene expression. Because the methylenetetrahydrofolate reductase (MTHFR) have had significant role in methionine metabolism polymorphisms into the gene could be related with early embryonic development. This study evaluated relationship between T allele in 677 C>T polymorphism in MTHFR and recurrent embryonic loss development. One hundred six women with tree or more pregnancy loss before 10 wg and 165 women without reproductive failure have been evaluated for 677 C>T carrier status. Sixteen (15.1%) of women with pregnancy loss have had TT genotype and 54 (50.9%) are heterozygous carriers for T allele. T allele frequency was higher but not significant differ from carrier status in control group (13.9% for TT genotype and 43.9% for CT OR and 95% CI respectively 1.1, 0.52-2.3 u 1.34, 0.8-2.26, p > 0.05). T allele (in homozygous and heterozygous carriers) was in higher but not significant prevalence in patients compared with controls (66% and 57.6% respectively, OR 1.43, 95% CI 0.84-2.46, p > 0.05), This study found a weak association between T allele carrier status (both in homozygous and heterozygous state) and recurrent embryonic loss development. T allele in 677 C>T polymorphism could be considered like an agent for early pregnancy wastage only in a constellation with other risk factors influencing embryonic development.

[DNA repair XRCC1, XPD genes polymorphism as associated with the development of bladder cancer and renal cell carcinoma].

We examined the correlations between the polymorphic alleles of the DNA repair genes XRCC1 (c.839G> A, rs25489; and c.1196A> G, rs25487), XPA (c.-4A> G, rs1800975), and XPD (c.2251A> C, rs13181) and the progression and severity of neoplasias in the bladder and kidney in patients of three distinct ethnic groups, Bashkir, Russians, and Tatar, residing in the Republic of Bashkorostan. The study enrolled 468 cancer patients and 351 healthy individuals. Genotyping for polymorphic alleles was carried out using the PCR-RFLP method. We identified a correlation between allele A of the c.839 G>A locus of the XRCC1 gene and the incidence of the bladder cancer (BC) and kidney cancer (KC) in the Tatar study group, using the additive genetic effects model (Odds Ratio (OR) = 5.23 and OR = 3.90). In turn, the heterozygous G/A genotype was present at a significantly higher frequency in the KC patients of Bashkir ethnic origin, compared with the control group (p = 0.0061, OR= 4.72). Additional analysis with consideration of participants' smoking status showed that the G/A genotype is significantly more frequent in smokers with BC (OR = 1.96, p = 0.05) then in healthy smokers. We also determined, using the recessive genetic model, that the genotype A/A of the c. 1196A>G locus of the XRCC1 gene was correlated with a higher risk of BC in the Russian cohort (OR = 2.29, p = 0.0082) and an increased incidence of KC in the Bashkir group (OR = 4.06, p = 0.05). A similar correlation was obtained for smokers. In contrast, the allele c.2251 A>C in the XPD gene correlated with a lower risk for BC and KC in the Tatars (p = 0.0003, OR = 0.48 and p < 0.0001, OR = 0.37) in the additive model and in the Bashkirs (p = 0.0083, OR = 0.12) and Russians (p = 0.0001, OR = 0.14) in the recessive model. Further, we uncovered that polymorphism c.839 G>A in the XRCC1 gene contributes to the progression of noninvasive and invasive BC and promotes KC at early and advanced stages of the disease. Thus, we identified similar correlations between DNA repair gene polymorphism and the incidence and progression of BC and KC. We propose that this result points to the involvement of common pathogenetic mechanisms in the initiation and progression of the urinary neoplasias.

[Results of open multicenter study of the safety of doxazosin in combination with indigal in men with stages I-II prostatic adenoma].

The article presents a method of conservative treatment of men with I-II stage prostatic adenoma using a combination of doxazosin and indigal, which has antioxidant, antiproliferative and anti-inflammatory properties, that allowed improving urodynamic parameters and reducing the progression prostate adenoma, minimizing the adverse effects of treatment.

Analysis of the efficiency of gene-cell therapy in transgenic mice with amyotrophic lateral sclerosis phenotype.

Amyotrophic lateral sclerosis is a neurodegenerative disease characterized by progressive death of cerebral and spinal motorneurons. Using behavioral tests we studied the efficiency of gene-cell therapy in SOD1 G93A transgenic mice receiving xenotransplantation of human umbilical cord blood mononuclear cells genetically modified with adenoviral vectors encoding vascular endothelial growth factor (VEGF) and reporter green fluorescent protein (EGFP) genes. The cells were transplanted to mice on week 27 of life (preclinical stage of the disease). Behavioral tests (open field, grip strength test) showed that transplantation of umbilical cord blood mononuclear cells expressing VEGF significantly improved the parameters of motor and explorative activity, grip strength, and animal survival. Thus, gene-cell therapy based on genetically modified mononuclear cells expressing VEGF can be efficient for the treatment of amyotrophic lateral sclerosis.

[Factor V Leiden in women with repeated IVF failures].

A plenty of factors have been connected with embryo implantation and further fetus development. Recurrent implantation failure (RIF) after assisted reproductive technology (ART) forces seeking the causes of decreased endometrial receptivity. A non-haemostatic function of thrombophilic mutations such as Factor V Leiden (FVL) was considered a factor related with endometrial receptivity. One hundred eighty eight women with two or more RIF after in vitro fertilization procedures investigated for carrier status for FVL was compared with carrier status of 97 women without reproductive failure who give a birth of at least one health child. There was no significant difference in carrier status for FVL in patients and controls (5.9% and 7.2% respectively, OR 0.80, 95% CI (0.26-2.73, p>0.05). Negligible higher prevalence of FVL was fond in health subjects compared with women with RIF A slightly positive relationship was found between FVL and embryo implantation. A preliminary determination of thrombophilic status in RIF women could specify needing or rejection of anticoagulant therapy during implantation period.

[Platelet intregrin beta3 A1/A2 polymorphism in women with stillbirth].

Maternal thrombophilia was recently discussed as possible cause for pregnancy complication, although the roles of some coagulation factors have not been clarified. Carrier status for platelet integrin beta3 polymorphism A1/A2 (PL A1/A2) was considered as possible risk factor for pregnancy complication. Seventy women with one or more stillbirth (intrauterine fetal death after 20 week of gestation) and 100 healthy control subjects were evaluated for PL A1/A2 to assess the impact of polymorphism for late pregnancy loss. The prevalence for PL A1/A2 in women with stillbirth was higher but not significantly differs from carrier status in control subjects (respectively 28.3% and 17%, OR = 1.93; 95% CI: 0.84 - 4.45). After adjustment for carrier status for Factor V Leiden (FVL) and Prothrombin (FII) gene mutation 20210 G > A the prevalence of PL 1/A2 remains a similar (28.2% O R = 1.92; 95% Il: 0.78 - 4.75). Combined carriers status for PL A1/ A2 with FVL or III 20210 G > A have had significantly higher prevalence in investigated group comparing with control subjects (respectively 20% and 2%, p < 0.0001). An independent impact of PL A1/A2 on risk of stillbirth development is not be yet established but additive role of the polymorphism in combination with other thrombophilic factors should be considered.

Accelerated photobleaching of a cyanine dye in the presence of a ternary target DNA, PNA probe, dye catalytic complex: a molecular diagnostic.

In many settings, molecular testing is needed but unavailable due to complexity and cost. Simple, rapid, and specific DNA detection technologies would provide important alternatives to existing detection methods. Here we report a novel, rapid nucleic acid detection method based on the accelerated photobleaching of the light-sensitive cyanine dye, 3,3'-diethylthiacarbocyanine iodide (DiSC(2)(3) I(-)), in the presence of a target genomic DNA and a complementary peptide nucleic acid (PNA) probe. On the basis of the UV-vis, circular dichroism, and fluorescence spectra of DiSC(2)(3) with PNA-DNA oligomer duplexes and on characterization of a product of photolysis of DiSC(2)(3) I(-), a possible reaction mechanism is proposed. We propose that (1) a novel complex forms between dye, PNA, and DNA, (2) this complex functions as a photosensitizer producing (1)O(2), and (3) the (1)O(2) produced promotes photobleaching of dye molecules in the mixture. Similar cyanine dyes (DiSC(3)(3), DiSC(4)(3), DiSC(5)(3), and DiSC(py)(3)) interact with preformed PNA-DNA oligomer duplexes but do not demonstrate an equivalent accelerated photobleaching effect in the presence of PNA and target genomic DNA. The feasibility of developing molecular diagnostic assays based on the accelerated photobleaching (the smartDNA assay) that results from the novel complex formed between DiSC(2)(3) and PNA-DNA is under way.

[Photodynamic therapy with visudyne: criteria for treatment success].

The paper specifies indications and success criteria for photodynamic therapy with visudine for classical macular choroidal neovascularization. Three major clinical types of a response to treatment are presented. These include: 1) complete regression of the subretinal neovascular membrane (SNM) with retinal edema resolution; 2) partial regression of SNM with its shift to inactive status that is attended by a significant reduction in pathological transudation from the newly formed vessels; 3) partial regression with the preserved high transudating activity of SNM. There was stabilization of lesion extents and visual functions in types 1 and 2 responses to therapy and a continued SNM growth and progressive visual worsening in type 3 response.

[New trends in surgical technologies of soft tissue wound management].

The authors performed an animal model study of postoperative wound complications using 72 white Wistar rats and 36 mongrel dogs with aseptic cut wounds, granulating wounds, and relaparotomic wounds. In the control group the wounds were closed by a conventional method; in the main group a machine stitch was performed. Wound closure in the main group was performed after approximation of wound edges with specially constructed devices. Morphological and microbiological analysis showed that the key role in development of complications is played by morphofunctional wound substrate, which comprises the degree of paravulnar tissue traumatization with suture when a conventional method is used, and paravulnar tissue ischemia in case of microbial dissemination. The authors conclude that machine stitch is preferable.

[Extrawound programmed vulnosynthesis in the treatment of festering wounds].

The treatment of 200 patients with granulating wound of soft tissues was analyzed. A conventional method of treatment was used in 90 patients. In the study group (110 patients) different types of adaptive-reposition devices and the method of extrawound programmed vulnosynthesis were used. This method demonstrated technical advantages compared with standard methods of treatment of festering wounds that permitted to improve the results of complex treatment.

[Experimental-clinical grounds for the treatment of granulating soft tissue wounds by the wound edge approximating stapler].

Factors of risk of the appearance of wound complications were studied in experiment on a model of granulating wounds in 18 mongrel dogs. A traditional method (control group) and an apparatus method (main group) were used for suturing the wounds. In the main group of animals suturing the wounds was performed after bringing together the wound edges by specially designed devices. Morphological and microbiological investigations have shown that the leading role in the appearance of complications belongs to the morphological substrate of the wound with the main component including the degree of traumatizing the paravulnerable tissue with a suture thread when connecting the wound edges by a traditional method, ischemia of the paravulnerable tissues under conditions of microbial dissemination. The introduction of the developed method of closing the soft tissue granulating wounds into clinical practice resulted in a statistically significant reduction of pyo-inflammatory complications in the main group of patients. A conclusion is made on a more favorable course of the wound process when using the apparatus method of closing the wounds.

[Surgical treatment of a carbuncle]. / Khirurgicheskoe lechenie karbunkula.

The results of the treatment of 250 patients with a carbuncle of various location were analyzed retrospectively. A differential approach to choice of surgical method with consideration of the degree, phase and localization of inflammation is preferable. Early and radical surgery, antimicrobic drugs, infusion therapy make base of up-to-date and adequate treatment. In the phase of infiltration deep incision often stops the disease or promotes a decrease of the necrotic zone. In suppurations linear incisions were made only on the face for cosmetic reasons. In other areas a few large connected T-, [symbol: see text]-, [symbol: see text]- or cross-formed incisions were preferable. In 30 cases I-formed incision with two rectangular flaps was used.

[Bougies for retrograde stretching of esophageal strictures]. / Buzhi dlia retrogradnogo rastiazheniia rubtsovykh struktur pishchedvoda.

The article deals with bougies (developed by its authors) of various types and sizes designed for gullet bougienage after chemical burns. The suggested bougies ensure less trauma to the esophagus walls during the bougienage procedure, which, in its turn, essentially cuts the time of recanalization and, according to authors' data, prevents ruptures in the internal tissues.