[Plasma miRNA expression in patients with genetically confirmed multiple endocrine neoplasia type 1 syndrome and its phenocopies].

BACKGROUND: MEN-1 is a rare autosomal dominant disease caused by mutations in MEN1 gene encoding the menin protein. This syndrome is characterized by the occurrence of parathyroid tumors, gastroenteropancreatic neuroendocrine tumors, pituitary adenomas, as well as other endocrine and non-endocrine tumors. If a patient with the MEN-1 phenotype carry no mutations in the MEN1 gene, the condition considers a phenocopy of syndrome (phMEN1). The possible cause of this changes could be changes in epigenetic regulation, particularly in microRNA expression that might affect menin signaling pathways. AIM: to identify differently expressed circulating miRNAs in plasma in patients with genetically confirmed MEN-1 syndrome, its phenocopies and healthy controls. MATERIALS AND METHODS: single-center, case-control study was conducted. We assessed plasma microRNA expression in patients with genetically confirmed MEN-1 (gMEN1), phMEN1 and healthy controls. Morning plasma samples were collected from fasting patients and stored at -80°C. Total RNA isolation was performed using miRNeasy Mini Kit with QIAcube. The libraries were prepared by the QIAseq miRNA Library Kit following the manufacturer. Circulating miRNA sequencing was done on Illumina NextSeq 500 (Illumina). Subsequent data processing was performed using the DESeq2 bioinformatics algorithm. RESULTS: we enrolled 21 consecutive patients with gMEN1 and 11 patients with phMEN1, along with 12 gender matched controls. Median age of gMEN1 was 38,0 [34,0; 41,0]; in phMEN1 - 59,0 [51,0; 60,0]; control - 59,5 [51,5; 62,5]. The gMEN1 group differed in age (p<0.01) but not gender (р=0.739) or BMI (р=0.116) compared to phMEN1 and controls group, the last two groups did not differ by these parameters (p>0.05). 25 microRNA were differently expressed in groups gMEN1 and phMEN1 (21 upregulated microRNAs, 4 - downregulated). Comparison of samples from the phMEN-1 group and relatively healthy controls revealed 10 differently expressed microRNAs: 5 - upregulated; 5 - downregulated. In the gMEN-1 and control groups, 26 differently expressed microRNAs were found: 24 - upregulated; 2 - downregulated. The miRNAs most differing in expression among the groups were selected for further validation by RT-qPCR (in the groups of gMEN1 vs phMEN1 - miR-3613-5p, miR-335-5p, miR-32-5p, miR-425-3p, miR-25-5p, miR-576-5p, miR-215-5p, miR-30a-3p, miR-141-3p, miR-760, miR-501-3p; gMEN1 vs control - miR-1976, miR-144-5p miR-532-3p, miR-375; as well as in phMEN1 vs control - miR-944, miR-191-5p, miR-98-5p). CONCLUSION: In a pilot study, we detected microRNAs that may be expressed differently between patients with gMEN-1 and phMEN-1. The results need to be validated using different measurement method with larger sample size.

[High tibial osteotomy with computer navigation for medial gonarthritis]. / Sravnitel'nye rezul'taty lecheniya medial'nogo gonartrita metodom vysokoi tibial'noi osteotomii s ispol'zovaniem komp'yuternoi navigatsii.

OBJECTIVE: To study significance of computer navigation for improving clinical and radiological results of medial gonarthritis treatment compared to non-invasive methods of lower limb axis correction control. MATERIAL AND METHODS: The study included 73 patients who were divided into 2 groups. The main group included 40 patients, the control group - 33 patients. In the main group, high tibial osteotomy was performed using computer navigation, in the control group - using non-invasive techniques. Clinical assessment was carried out according to the KSS, KOOS and VAS scales. We assessed the main reference angles of the lower limb considering X-ray data. RESULTS: Both groups were characterized by postoperative improvement of clinical results according to various scales. Computer navigation provided higher accuracy in most cases. We focused on target correction of 3° valgus. CONCLUSION: High tibial osteotomy with computer navigation or non-invasive techniques is an effective treatment method for medial gonarthritis. There are no significant differences in clinical results according to the KSS and KOOS scales, as well as X-ray data after correction. We found significant differences in VAS scores.

[INVESTIGATION OF SURGICAL CORRECTION OF ANTERIOR APICAL PROLAPSE OF THE PELVIC ORGANS USING A COMBINED TECHNIQUE, MONOLATERAL CYRENE POSTERIOR IN COMBINATION WITH ANTERIOR COLPORRHAPHY].

Objective - to evaluate the results of surgical correction of anterior apical prolapse of the pelvic organs with a combined technique monolateral CYRENE posterior (using monofilament polypropylene tape CYRENE) in combination with anterior colporrhaphy. A prospective cohort study included 240 patients with urogenital prolapse. The subjects were followed up for three months, and the anatomical outcome according to the POP-Q System classification, intra- and postoperative complications, absence or presence of relapse, quality of life and sex life were assessed. A relapse was considered to be grade 3 or more anterior prolapse, grade 2 or more central prolapse. Quality of life and sex life were assessed using the PFDI-20, PFIQ-7 and PSIQ-12 questionnaires. The average age of women was 57.8 years. All 240 patients had grade 3 cystocele, prolapse of the anterior vaginal wall, and prolapse of the uterus. In 23 (9.6%) patients, de novo urinary incontinence developed, 8 patients had stress urinary incontinence before surgery, 6 of them persisted stress urinary incontinence after surgery, 2 of them stress urinary incontinence was arrested without the simultaneous use of a mid-urethral sling. According to questionnaires, the quality of life improved in 225 (96.9%) women according to PFDI-20 and in 210 (90.5%) according to PFIQ-7. There were no changes in the quality of sex life. Anatomically, 235 (96.9%) achieved the desired result (Ba≤ II, C≤ I according to the POPQ System). At the same time, in 36 (15%), 3 months after the operation, cystocele of the 2nd degree was revealed. In all cases, there was minimal blood loss, in 9 cases (3.75%) there was a hematoma of the anterior vaginal wall that did not require additional intervention, at discharge had signs of organization and was completely stopped 6 weeks after surgery; 3 (1.25%) patients had urinary retention after surgery. Dyspareunia was not identified in any patient. The study of the treatment of urogenital prolapse using a combined technique, monolateral CYRENE posterior in combination with anterior colporrhaphy, demonstrates high efficiency and relative safety. The technique showed excellent anatomical results for apical prolapse and good for anterior prolapse. A small number and low significance of postoperative complications were revealed. Longer postoperative follow-up is needed to assess long-term results and long-term prospects in order to form a more complete understanding of this technique.

[Clinical and functional outcomes of valgus osteotomy in patients with medial gonarthrosis (preliminary results)]. / Rezul'taty val'giziruyushchei osteotomii u patsientov s medial'nym gonartrozom (predvaritel'nye rezul'taty).

OBJECTIVE: To analyze gait biomechanics before and after corrective osteotomies around the knee joint in patients with medial gonarthrosis. MATERIAL AND METHODS: The study recruited 5 patients (6 surgeries) with medial gonartrosis and varus knee deformity. Gait biomechanics was analyzed in preoperative period and 6 months later. The control group consisted of 20 healthy people. RESULTS: Knee joint biomechanics was almost normal in postoperative period. Amplitude of knee joint flexion was increased. Moreover, hip joint flexion-extension amplitude was increased on both sides and often exceeded normal values. Clinical results showed significant increase in IKS (from 55 to 89.6) and VAS score (from 7.7 to 2.3). Postoperative correction angle varied within 1.5-2° of valgus. CONCLUSION: Corrective osteotomy around the knee joint is effective for medial gonarthrosis. We obtained favorable clinical results and limb axis correction after surgery without significant hypercorrection. Gait biomechanics was also improved.

[Short-term and long-term remission after endoscopic transnasal adenomectomy in patients with acromegaly].

BACKGROUND: Neurosurgery is the most effective treatment for acromegaly. As most of the patients present with macroadenomas, surgical treatment is not always successful, even with the expert level of a neurosurgeon. Assessment of the postoperative remission rates in acromegaly preoperative predictors of treatment efficacy is an urgent task of modern research. AIM: To assess the short-term and long-term remission of acromegaly after endoscopic transnasal adenomectomy in a tertiary medical center and assess preoperative predictors of the treatment effectiveness. MATERIALS AND METHODS: A single-center, prospective, uncontrolled study was conducted. We included patients with active acromegaly who did not receive medical therapy with somatostatin analogues and were referred for endoscopic transsphenoidal adenomectomy. Plasma miRNA expression was assessed by quantitative reverse transcription PCR. Postoperative samples of adenomas were sent for study, with the determination of the immunohistochemical staining for somatostatin receptors 2 and 5 subtypes and morphology was performed on postoperative adenoma samples. RESULTS: The study included 44 patients: 32.8% men, median age 47.0 [34.0; 55.0], IGF-1 744.75 ng/ml [548.83;889.85], growth hormone 9.5 ng/ml [4.94; 17.07]. Tumor volume 832 mm3 [419.25; 2532.38]. Early postoperative remission was achieved in 35 patients (79.5%). Patients who achieved short-term remission had higher IGF-1 and basal growth hormone levels. Median follow-up was 19.0 months [12.5;29.0]. Long-term remission was achieved in 61.4% (27 patients), no remission in 9 (20.5%), recurrency in 2 patients (4.5%), 6 patients were to follow-up (13.6%). In patients with long-term remission, we observed lower growth hormone and IGF-1 levels. No differences in miRNA expression was observesd. The predictive value of basal GH before surgery for long-term remission was assessed: area under the curve 0.811 (95% CI: 0.649; 0.973). A cut-off value of 15.55 ng/mL corresponded to a sensitivity of 70.0% (34.8%; 93.3%), a specificity of 85.7% (67.3%; 96.0%), an accuracy of 81.6% (65 .7%; 92.3%), PPV 63.6% (39.3%; 82.5%), NPV 88.9% (75.4%; 95.4%). CONCLUSION: Rates of short-term and long-term remission after endoscopic transsphenoidal adenomectomy in our cohort is 79,5% и 61,4%, respectively, and is comparable with literature data for expert pituitary centers. Preoperative GH shows potential value in predicting the long-term remission of acromegaly, but further studies in a larger sample are needed to obtain more accurate cut-off values.

The value of indicators characterizing the state of the cardiovascular system in assessing the hospital prognosis of COVID-19 patients.

Background     Heart damage is one of complications of the novel coronavirus infection. Searching for available predictors for in-hospital death and survival that determine the tactic of managing patients with COVID-19, is a challenge of the present time.Aim      To determine the role echocardiographic (EchoCG) parameters in evaluation of the in-hospital prognosis for patients with the novel coronavirus infection, COVID-19.Material and methods  The study included 158 patients admitted for COVID-19. EchoCG was performed for all patients. The role of left ventricular (LV) ejection fraction (EF) was analyzed in various age groups. EchoCG data were compared with the clinical picture, including the severity of respiratory failure (RF), blood oxygen saturation (SрО2), data of computed tomography (CT) of the lungs, and blood concentration of troponin. Comorbidity was analyzed, and the highest significance of individual pathologies was determined.Results LV EF ≤40 % determined the worst prognosis of patients with COVID-19 (p<0.0001), including the age group older than 70 years (р=0.013). LV EF did not correlate with the degree of lung tissue damage determined by CT upon admission (р=0.54) and over time (р=0.23). The indexes that determined an adverse in-hospital prognosis to a considerable degree were pericardial effusion (p<0.0001) and pulmonary hypertension (p<0.0001). RV end-diastolic dimension and LV end-diastolic volume did not determine the in-hospital mortality and survival. Blood serum concentration of troponin I higher than 165.13 µg/l was an important predictor for in-hospital death with a high degree of significance (р<0.0001). Th degree of RF considerably influenced the in-hospital mortality (р<0.0001). RF severity was associated with LV EF (р=0.024). The SpO2 value determined an adverse immediate prognosis with a high degree of significance (р=0.0009). This parameter weakly correlated with LV EF (r=0.26; p=0.0009). Patients who required artificial ventilation (AV) constituted a group with the worst survival rate (р<0.0001). LV EF was associated with a need for AV with a high degree of significance (р=0.0006). Comorbidities, such as chronic kidney disease, postinfarction cardiosclerosis and oncologic diseases, to the greatest extent determined the risk of fatal outcome.Conclusion      EchoCG can be recommended for patients with COVID-19 at the hospital stage to determine the tactics of management and for the in-hospital prognosis.

[The Influence of Adiponectin on Production of Apolipoproteins A-l and E by Human Macrophages].

Adiponectin is an adipose tissue hormone affecting energy and lipoprotein metabolism and modulating inflammatory responses. However, the role of this adipokine in atherogenesis remains poorly understood. The aim of this study was to investigate the effect of adiponectin on the production of apolipoproteins (apo) A-l and E by human macrophages (MP). The study was conducted on macrophage-like cells of the THP-1 cell line of two differentiation terms, 3 and 5 days (3d and 5d). Adiponectin (10 µg/mL) stimulated the expression of apoA-1 gene at the mRNA level in 5d MP, but not in 3d MP. The level of apoE mRNA in MP under the action of adiponectin was not affected. Adiponectin suppressed macrophage TNF gene expression, while it induced the expression of IL-10 gene in 5d MP. The secreted levels of apoA-1 and apoE proteins under the action of adiponectin in macrophages of both periods of differentiation remained unchanged, while the level of the surface apoA-1 protein in 5d MP was decreasing. Incubation of 5d MP with the PPARα nuclear receptor antagonist MK-886 or with the nuclear receptor LXR agonist TO-901317 resulted in cancellation of the stimulating effect of adiponectin on apoA-1 gene expression. These data indicate that adiponectin, in addition to its anti-inflammatory action, has a modulating effect on production of apoA-1 by macrophages. The latter is probably one of the mechanisms of the influence of this adipokine on atherogenesis.

Non-magnetic shell coating of magnetic nanoparticles as key factor of toxicity for cancer cells in a low frequency alternating magnetic field.

This work is devoted to studying the effects of non-magnetic shell coating on nanoparticles in a low frequency alternating magnetic field (LF AMF) on tumor cells in vitro. Two types of iron oxide nanoparticles with the same magnetic core with and without silica shells were synthesized. Nanoparticles with silica shells significantly decreased the viability of PC3 cancer cells in a low frequency alternating magnetic field according to the cytotoxicity test, unlike uncoated nanoparticles. We showed that cell death results from the intracellular membrane integrity failure, and the calcium ions concentration increase with the subsequent necrosis. Transmission electron microscopy images showed that the uncoated silica nanoparticles are primarily found in an aggregated form in cells. We believe that uncoated nanoparticles lose their colloidal stability in an acidic endosomal environment after internalization into the cell due to surface etching and the formation of aggregates. As a result, they encounter high endosomal macromolecular viscosity and become unable to rotate efficiently. We assume that effective rotation of nanoparticles causes cell death. In turn, silica shell coating increases nanoparticles stability, preventing aggregation in endosomes. Thus, we propose that the colloidal stability of magnetic nanoparticles inside cells is one of the key factors for effective magneto-mechanical actuation.

Multiple bilateral hip fractures in a patient with dyskeratosis congenita caused by a novel mutation in the PARN gene.

We report a case of a young male patient with clinical signs of dyskeratosis congenita who presented with multiple bilateral low-traumatic hip fractures. Whole exome sequencing (WES) showed a previously unreported mutation in the poly(A)-specific ribonuclease (PARN) gene. Zoledronic acid 5 mg over 3 years was effective at preventing further fractures. A male patient was referred to our clinic at age 24 due to multiple bilateral hip fractures. At the time of admission, the patient's height was 160 cm and weight 40 kg; bone mineral density (BMD) at the lumbar spine was normal (L1-L4 0.0 Z-score). The patient was found to have abnormal skin pigmentation, hyperkeratosis of palms and soles, nail dystrophy, and signs of bone marrow failure (BMF). Bone fragility first presented at 5 years old with a wrist fracture, followed by multiple bilateral low-traumatic hip fractures without falls from 14 to 24 years. WES showed a previously unreported mutation (NM_002582.3: c.1652delA; p.His551fs) in the poly(A)-specific ribonuclease (PARN) gene. Flow fish telomere measurement result was 5.9 (reference range 8.0-12.6), which is consistent with the DC diagnosis. Permanent fixation with internal metal rods and zoledronic acid 5 mg over 3 years was effective at preventing further fractures over 4 years of follow-up. Additionally, BMF did not progress over 4 years of observation. DC associated with PARN gene mutations might predispose to low-traumatic multiple hip fractures in adolescents and young adults. Treatment with zoledronic acid in this case was effective and safe at preventing further fractures.

Results of in vitro test-KRAStissue reagent kit test to determine indications for target therapy of patients diagnosed with colorectal cancer.

As subjects of the clinical trial, 44 samples of paraffin-fixed tissue were used from patients diagnosed with "colorectal cancer." In the course of clinical trials, 44 samples of paraffin-fixed tissue were analyzed in two series of experiments, that is, 88 clinical-laboratory experiments were carried out, of which 48 experiments with genomic DNA samples with the established negative status of the presence of KRAS gene mutations and 40 experiments with genomic DNA samples with the established positive status of the presence of KRAS gene mutations. Analysis and evaluation of the results of clinical laboratory tests of the medical product "Kit of Reagents for Determination of the Status of KRAS Gene Mutations by PCR-RV Method in a Sample of Human Genomic DNA from Samples of Paraffin-Fixed Tissue (Test-KRAS-tissue) according to TU 21.20.23-006-97638376-2016 "confirmed that it allows to carry out qualitative determination of the status of six mutations of the twelfth codon (Gly12Asp, Gly12Ala, Gly12Arg, Gly12Val, Gly12Ser, Gly12Cys) and one mutation of the thirteenth codon (Gly13Asp) the KRAS gene by real-time allele-specific PCR in human genomic DNA sample from paraffin-fixed tissue samples, with high diagnostic sensitivity rates of 90.9% and diagnostic specificity of 95.0% with a confidence probability of 90%. Reproducibility of results is 100%, which confirms the high reliability of the set.

[Rigid transpedicular fusion surgery in the treatment of degenerative lumbar stenosis]. / Rigidnaya transpedikulyarnaya fiksatsiya v lechenii patsientov s degenerativnym poyasnichnym stenozom.

OBJECTIVE: To compare an effectiveness of different methods of rigid transpedicular fixation and decompression in patients with degenerative lumbar stenosis. MATERIAL AND METHODS: A prospective study included 160 patients with degenerative lumbar spine stenosis. In the first group (n=37), patients underwent laminectomy and transpedicular fixation, in the second group (n=60) - laminectomy, transpedicular fixation and implantation of interbody fusion cage. In the third group (n=30), interlaminar decompression and transpedicular fixation were carried oud, in the fourth group (n=33) - interlaminar decompression, transpedicular fixation and implantation of interbody fusion cage. Surgeries were performed in three clinics in Moscow. Outcomes were assessed using a visual analogue scale (VAS) and Oswestry questionnaire in 1 and 2 years after surgery. Between-group comparison of the outcomes was performed. In patients with unfavorable outcome, we analyzed the cause of unsatisfactory result and risk factors. RESULTS: Satisfactory result was noted in 103 patients (64%) in 2 years after surgery. Outcomes were comparable in all groups. More significant regression of back pain was noted in group II (laminectomy, transpedicular fixation, interbody cage) compared to other groups. Preoperative risk factors of adverse outcome were resting leg pain VAS score > 4 and age over 71 years. Incidence of pseudoarthrosis and back pain was higher among patients without interbody cage. Incidence of adjacent level lesion was higher among patients with interbody cages. CONCLUSION: Decompression type and implantation of interbody cage do not significantly change postoperative outcomes in most patients with degenerative lumbar stenosis undergoing transpedicular fixation. However, interbody cage implantation during transpedicular fixation is advisable in patients with severe back pain (VAS score > 5-6).

[TOTRP EXPERIENCE IN TREATING WOMEN'S STRESS CONTINUITY IN WOMEN].

The aim of the study was to evaluate the efficacy and safety of the TOTRP (tension-free vaginal tape-obturator retropublic) methodology, as well as the frequency of de novo urgency symptoms. A retrospective study included 32 women with stressful and mixed urinary incontinence with a predominance of the stress component without concomitant genital prolapse, as well as without surgical intervention for a history of urinary incontinence and pelvic prolapse. Patients underwent surgical intervention according to the TOTRP method using Lintex UroSling® polypropylene tape from February to October 2018. According to the results of the ICIQ-SF and OAB awareness Tool questionnaires, 14 (43.7%) patients were registered prior to surgical treatment. with an average degree of stress urinary incontinence, 18 (56.3%) - severe; 19 (59%) patients with mild bladder hyperactivity, 4 (12.5%) - moderate. Through surgical intervention, stress urinary incontinence was completely stopped in 26 (81.25%) and transferred to mild degree in 2 (6.25%), in 5 (15.6%) - moderate degree. So, the effectiveness of the TOTRP technique was 87.5%. Postoperative urgency was 8 (25%), of which de novo - 5 (15.63%). The treatment was characterized by the complete absence of intraoperative complications.

[Malignant cerebellar infarction: clinical course and surgical treatment]. / Zlokachestvennyi infarkt mozzhechka: klinicheskoe techenie i khirurgicheskoe lechenie.

AIM: To clarify the indications for surgical treatment of malignant cerebellar infarction (CI). MATERIAL AND METHODS: Eighty patients with CI were studied. The malignancy of CI was understood as the development of mass effect in the posterior cranial fossa, accompanied by the decrease in consciousness due to compression of the brain stem and/or the development of occlusive hydrocephalus. The patients were divided into 2 groups. The group of malignant CI included 55 patients (68.75%) (group I), the group of benign CI included 25 patients (31.25%) (group II). Patients of group I were divided into subgroups, one of them underwent surgical treatment (surgical subgroup), and another only conservative (conservative subgroup) treatment. Surgery efficacy criteria were: restoration of consciousness to 15 points according to GCS and/or restoration of the fourth ventricle and the quadrigeminal cistern configurations. Results of treatment were assessed according to the Glasgow outcome scale. RESULTS: Malignant CI occurred more frequently in patients with the volume of ischemia exceeding 20 cm3 (p<0.05) in the first day of the disease. The threshold value of mass effect, which can cause further a malignant CI, was 3 points according to the M. Jauss scale. In the group of patients with malignant CI, surgical treatment reduced the mortality rate from occlusion and dislocation syndrome by 35.8%. The most effective type of intervention was a combination of decompressive trepanation of the posterior cranial fossa and external ventricular drainage. CONCLUSION: In patients with CI with the volume more than 20 cm3 and signs of mass effect in the posterior cranial fossa on the scale of M. Jauss 3 points or more, the malignant course of the disease develops in 67% of cases. These patients require careful monitoring, and, in case of development of malignant CI, surgical treatment is necessary.

Role of MicroRNAs in the Regulation of Subcutaneous White Adipose Tissue in Individuals With Obesity and Without Type 2 Diabetes.

Obesity is a high-risk factor for such comorbidities as cardiovascular disease, several types of cancer, and type 2 diabetes; however not all individuals with obesity have such complications. Approximately 20% of individuals with obesity are metabolically healthy. This study focused on differences between obese individuals with and without type 2 diabetes (T2D+ and T2D-, respectively) on the transcriptome level. Subjects included were 35 T2D- patients with obesity and 35 T2D+ patients with obesity with the same body mass index (BMI). The study was based on the transcription analysis of mRNA and microRNAs (miRs) by RNAseq. In the first step, we performed RNAseq of miRs, in the second step, we analyzed only those mRNA, which appeared targets for significant miRs from the first step. All RNAseq results were validated by qPCR. There were seven miRs differently expressed with adjusted p-value <0.1, which were confirmed by qPCR. Five among them: miR-204-5p, miR125b-5p, miR-125a-5p, miR320a, miR-99b-were upregulated in T2D+ patients with obesity, while only two miRs, miR-23b-3p, and miR197-3p, were increased in T2D- patients with obesity. These seven miRs target two groups of genes: matrix metalloproteinases and TGFß signal pathway genes. According to the results of transcriptome analysis, the main difference between T2D+ and T2D- patients with obesity was in adipogenesis and fibrosis regulation by matrix metalloproteinases and SMAD4-RUNX2 signal cascade. Based on the data about transcription profiles of both groups, we suggested that the process of fibrosis in T2D+ patients with obesity is more pronounced than in T2D- patients with obesity.

Chemical Reprogramming of Somatic Cells in Neural Direction: Myth or Reality?

In in vitro experiments on cultures of human multipotent stem cells from the human bonearrow and dental pulp, we studied direct reprogramming towards neuro-glial lineage cells using a cocktail of small molecules. Reprogramming by the previously published protocol (with a cocktail containing ß-mercaptoethanol, LIF, VPA, CHIR99021, and RepSox) and by the optimized protocol (VPA, RG108, А83-01, dorsomorphin, thiazovivin, CHIR99021, forskolin, and Isx9) allows obtaining cells with immunophenotypic and genetic signs of neural stem cells. However, neither the former, nor the optimized protocols allowed preparing neural progenitors capable of adequate terminal differentiation from both bone marrow-derived mesenchymal stem cells and nestin-positive neural crest-derived mesenchymal stem cells. Real-time PCR demonstrated the expression of some neurogenesis markers, but neural stem cell-specific expression pattern was not observed. The findings lead us to a conclusion that reprogramming with small molecules without additional factors modifying gene expression does not allow reproducible production of human neural stem cell-like progenitors that can be used as the source of neural tissue for the regenerative therapy.

[Perspectives of improvement of the diagnosis of prostate cancer based on analysis of PCA3 gene expression].

Prostate cancer is the second leading cause of cancer death. The widespread introduction into the clinical practice of test for prostate specific antigen (PSA) led to an increase in the number of prostate biopsies performed. At the same time, a decrease in the threshold of age-specific PSA standards has resulted in an increase in the number of unnecessary biopsies. In this regard, a need has arisen for new prostate cancer biomarkers. PCA3 is a non-coding mRNA that is exclusively expressed by prostate cells. Currently, three generations of test diagnostic systems based on the quantitative analysis of the PCA3 mRNA in the urine or its cell sediment has already developed, and they differ in the type of material studied and the method for estimating the amount of PCA3 mRNA. Clinical studies of the developed test systems have shown that a high level of PCA3 expression in the patients urine correlates with the probability of detecting prostate cancer. PCA3 test has higher positive and negative predictive values than previously used PSA test. These data are repeatedly confirmed by studies conducted in different clinics. Thus, the introduction of the method of quantitative determination of PCA3 in clinical practice can significantly improve the efficiency of diagnosis of prostate cancer and reduce the number of unnecessary biopsies.

Hypoxia as a Factor Involved in the Regulation of the apoA-1, ABCA1, and Complement C3 Gene Expression in Human Macrophages.

Hypoxia plays a critical role in progression of atherosclerosis. Local oxygen deficiency in a plaque creates a specific microenvironment that alters the transcriptome of resident cells, particularly of macrophages. Reverse cholesterol transport from plaque to liver is considered a main mechanism for regression of atherosclerosis. Ubiquitously expressed ATP-binding cassette transporter A1 (ABCA1) and liver- and small intestine-derived apolipoprotein A-1 (ApoA-1) are two main actors in this process. We recently reported endogenous apoA-1 expression in human macrophages. While ABCA1 and ApoA-1 have antiatherogenic properties, the role of complement factor C3 is controversial. Plasma C3 level positively correlates with the risk of cardiovascular diseases. On the other hand, C3 gene knockout in a murine atherosclerosis model increases both plaque size and triglycerides level in blood. In the present study, we show for the first time that a hypoxia-mimicking agent, CoCl2, induces the upregulation of the apoA-1 and C3 genes and the accumulation of intracellular and membrane protein ApoA-1 in THP-1 macrophages. The MEK1/2-Erk1/2 and MKK4/7-JNK1/2/3 cascades are involved in upregulation of ABCA1 and C3 via activation of transcription factor NF-κB, which interacts with the HIF-1α subunit of hypoxia-inducible factor 1 (HIF-1). The three major MAP-kinase cascades (Erk1/2, JNK1/2/3, and p38) and the NF-κB transcription factor are involved in the hypoxia-induced expression of the apoA-1 gene in THP-1 macrophages.

[Expression of plasma microRNA in patients with acromegaly].

BACKGROUND: microRNA is a class of small non-coding RNA molecules involved in posttranscriptional regulation of gene expression. MicroRNAs are detectable in blood in stable concentrations, which makes them promising biomarkers for various diseases. AIM: to assess plasma microRNA expression in patients with active acromegaly compared with healthy controls. MATERIAL AND METHODS: single-center, case-control study: assessment of plasma microRNA in patients with acromegaly compared with healthy controls. Fasting blood samples were drawn and centrifuged at +5С temperature and 3000 rpm for 20 minutes, then aliquoted and frozen at 80C until further analysis. MicroRNA extraction and library preparation was done according to manufacturers instructions. Expression analysis was performed on NextSeq sequencer. Bioinformatic analysis using atropos (adapted deletion), STAR (aligning), FastQC (quality control), seqbuster/seqcluster/miRge2 (microRNA annotation, isomiR and new microRNA search, expression analysis). Primary endpoint of the study differential expression of plasma microRNA in patients with acromegaly compared with healthy controls. RESULTS: we included 12 patients with acromegaly age 33.1 [20; 47], BMI 29.3 kg/m2 [24.0; 39.6], IGF-1 686.1 ng/mL [405.9; 1186.0] and 12 healthy subjects age 36.2 [26; 44], BMI 26.7 kg/m2 [19.5; 42.5], IGF-1 210.4 ng/mL [89.76; 281.90]; gender ratio for both groups 4 males, 8 females. The groups did not differ in gender (p=0.666), age (p=0.551) and BMI (p=0.378). We found decreased expression of four microRNAs in patients with acromegaly: miR-4446-3p 1.317 (p=0.001), miR-215-5p 3.040 (p=0.005), miR-342-5p 1.875 (p=0.013) and miR-191-5p 0.549 (p=0.039). However, none of these changes were statistically significant after adjustment for multiple comparisons (q 0.1). CONCLUSION: we found four microRNAs, which could potentially be downregulated in plasma of patients with acromegaly. The result need to be validated using different measurement method with larger sample size.

Biodistribution and Tumors MRI Contrast Enhancement of Magnetic Nanocubes, Nanoclusters, and Nanorods in Multiple Mice Models.

Magnetic resonance imaging (MRI) is a powerful technique for tumor diagnostics. Iron oxide nanoparticles (IONPs) are safe and biocompatible tools that can be used for further enhancing MR tumor contrasting. Although numerous IONPs have been proposed as MRI contrast agents, low delivery rates to tumor site limit its application. IONPs accumulation in malignancies depends on both IONPs characteristics and tumor properties. In the current paper, three differently shaped Pluronic F-127-modified IONPs (nanocubes, nanoclusters, and nanorods) were compared side by side in three murine tumor models (4T1 breast cancer, B16 melanoma, and CT26 colon cancer). Orthotopic B16 tumors demonstrated more efficient IONPs uptake than heterotopic implants. Magnetic nanocubes (MNCb) had the highest r2-relaxivity in vitro (300 mM-1·s-1) compared with magnetic nanoclusters (MNCl, 104 mM-1·s-1) and magnetic nanorods (MNRd, 51 mM-1·s-1). As measured by atomic emission spectroscopy, MNCb also demonstrated better delivery efficiency to tumors (3.79% ID) than MNCl (2.94% ID) and MNRd (1.21% ID). Nevertheless, MNCl overperformed its counterparts in tumor imaging, providing contrast enhancement in 96% of studied malignancies, whereas MNCb and MNRd were detected by MRI in 73% and 63% of tumors, respectively. Maximum MR contrasting efficiency for MNCb and MNCl was around 6-24 hours after systemic administration, whereas for MNRd maximum contrast enhancement was found within first 30 minutes upon treatment. Presumably, MNRd poor MRI performance was due to low r2-relaxivity and rapid clearance by lungs (17.3% ID) immediately after injection. MNCb and MNCl were mainly captured by the liver and spleen without significant accumulation in the lungs, kidneys, and heart. High biocompatibility and profound accumulation in tumor tissues make MNCb and MNCl the promising platforms for MRI-based tumor diagnostics and drug delivery.