[Comparative analysis of clinical features of robotic-assisted and laparoscopic partial nephrectomy].

INTRODUCTION: Kidney cancer (mostly renal cell carcinoma) is one of the ten most commonly diagnosed malignant tumors among men and women. Due to the widespread use of computer tomography and magnetic resonance imaging, the proportion of early-stage kidney cancers has increased. Currently, treatment options for stage 1 kidney cancer are radical nephrectomy, partial nephrectomy, and active surveillance. Among organ-preserving intervention, three main techniques can be distinguished: open surgery, minimally invasive surgery and ablation methods. To date, robotic-assisted procedures have occupied their place among minimally invasive interventions. AIM: To carry out a comparative analysis of two methods of organ-preserving treatment of kidney tumors, namely robot-assisted and laparoscopic partial nephrectomy. MATERIALS AND METHODS: A retrospective comparative analysis of two groups of patients with kidney tumors who underwent robotic-assisted or laparoscopic partial nephrectomy during the period from 2012 to 2019 was performed. RESULTS: There were no differences between two groups in age, mean score on the RENAL nephrometry scale, preoperative creatinine levels, tumor size, and duration of warm ischemia. However, duration of surgery, the volume of blood loss, serum creatinine after surgery, the length of stay, the use of the technique of early unclamping of the renal artery, the use of technique "off-clamp" and the proportion of exophytic tumors with growth were significantly different between patients of two groups. CONCLUSION: We believe that the robotic system is intuitively convenient for performing partial nephrectomy, allowing the treatment of potentially more complex cases and expanding the indications for organ-preserving procedures.

[Robot-assisted laparoscopic partial nephrectomy].

Currently, partial nephrectomy as a nephron-sparing surgery is the standard treatment method of malignant kidney tumors stage T1 in the absence of contraindications. Robotic system is an intuitively convenient platform for performing partial nephrectomy of various degrees of complexity. The technique of robot-assisted laparoscopic partial nephrectomy continues to improve but is still not standardized. A description of surgical technique, results and complications of robot-assisted laparoscopic partial nephrectomy are presented in the article.

[Fibrinolytics: from the thrombolysis to the processes of blood vessels growth and remodeling, neurogenesis, carcinogenesis and fibrosis].

One of the most outstanding scientific achievements in the thrombolysis is the development and administration of fibrinolysin - the first Soviet drug that lyses blood clots. Intracoronary administration of fibrinolysin reduced the mortality of patients with myocardial infarction by almost 20%. For his work in this field Yevgeny Chazov was awarded the Lenin Prize in 1982. Over the next decades, under his leadership, the Cardiology Center established scientific and clinical laboratories that created new generations of drugs based on fibrinolytics for treating patients with myocardial infarction, restoration of blood flow in ischemic tissue, and also studying the mechanisms of remodeling of blood vessels involving the fibrinolysis system. It have been found new mechanisms of regulation of the navigation of blood vessels and nerves growth, tumor growth and its metastasis with the participation of the fibrinolysis system proteins. The review reports the role of the fibrinolysis system in the thrombolysis, blood vessels growth and remodeling, neurogenesis, carcinogenesis and fibrosis. The article is dedicated to the 90th anniversary of academician E.I. Chazov.

[THE METHODICAL APPROACHES TO DIAGNOSTIC OF NIGHT PAROXYSMAL HEMOGLOBINURIA].

The article presents diagnostic of night paroxysmal hemoglobinuria. The night paroxysmal hemoglobinuria is an orphan disease characterized by absence of GPI-anchor on blood cells as a result of mutation of PIG-A gene on the short arm of X-chromosome. The particular proteins bounded with GPI-anchor implement function of defense from activation of components of complement and development of membrane-attacking complex. The erythrocytes exposed to destruction in bloodstream are among the most impacted. Therefore, one of the main signs of night paroxysmal hemoglobinuria is complement-depending intravascular hemolysis which indicators for a long time played a key role in diagnostic of night paroxysmal hemoglobinuria. The actual technique of diagnostic of night paroxysmal hemoglobinuria is flow cytometry. The analysis of night paroxysmal hemoglobinuria clone is recommended to patients with hemolysis of unclear genesis, thrombosis of cerebral and abdominal veins, thrombocytopenia and macrocytosis and also patients with AA, myelodysplastic syndrome, myelofibrosis. The international protocol recommended by the International Society of Clinical Cytometry (2010) is implemented to diagnose night paroxysmal hemoglobinuria. The original technique of evaluation of reticulocytes was developed with purpose to detect night paroxysmal hemoglobinuria clone. The high correlation was substantiated between size of night paroxysmal hemoglobinuria clone measured among reticulocytes according to proposed mode and night paroxysmal hemoglobinuria clone measured among granulocytes and monocytes detected according international standardized approach.

[Mechanisms of Vascular Remodeling Following Arterial Injury].

Arterial remodeling is the process of adaptation of the vessel comprising multiple structural and functional alterations of the vascular wall that occur in disease, trauma or aging. Arterial remodeling is accelerated in conditions that adversely affect the structural and functional balance of the vascular system, such as hypertension, atherosclerosis, kidney disease, inflammatory diseases, genetic abnormalities, and mechanical damage. Pathological changes in the vascular wall lead to organ damage and, ultimately, death. The aim of this paper is to review the various factors and complex mechanisms, which underlie negative arterial remodeling after mechanical injury and data indicating on the key role of the urokinase plasminogen activator in this process.

Non-viral transfer of BDNF and uPA stimulates peripheral nerve regeneration.

Peripheral nerves connect brain and spinal cord with the extremities and inner organs, and nerves injury can lead the disability and social exclusion. Growth factors and other natural stimulators of regeneration processes look very promising as future medicines. In our study, we tested the influence of genetic constructions that contain genes of brain-derived neurotrophic factor and urokinase plasminogen activator on nerve's structure and function after traumatic and ischemic injuries. Injection of pVax1-hBDNF and pVax1-muPA after traumatic injury led to better restoration of nerve's structure and function compared to similar parameters of control group mice. In ischemic injury model pVax1-hBDNF and pVax1-muPA slowed and reduced the damage progression and stimulated nerve regeneration as well. However, the treatment with pVax1-muPA was less effective after the traumatic injury. As we chose a non-viral method of gene delivery during our study the optimal conditions of plasmid intramuscular delivery were also determined.

Molecular modeling as a new approach to the development of urokinase inhibitors.

Proteolytic activity of urokinase plays an important role in negative remodeling of blood vessels, restenosis, tumor angiogenesis, and metastasizing, which necessitates the development of selective urokinase inhibitors. Using methods of computer modeling (docking, post processing, and direct docking) and quantum chemistry, we selected substances from the large compound database, analyzed their structures, and experimentally verified their inhibitor activity. New urokinase inhibitor candidates were proposed based on the theoretical predictions and experimental verification of compound activities. The process of modifying urokinase inhibitors based on (benzothiazol-3-yl)guanidine was developed. A new urokinase inhibitor (5-brom-benzothiazol-3-yl)guanidine, that can be effective for regulation of vascular remodeling and tumor angiogenesis, was created.

[The evaluation and comparative characteristic of detection of clone of paroxysmal nocturnal hemoglobinuria on reticulocytes using the technique of flow cytometry].

The paroxysmal nocturnal hemoglobinuria is a rare clonal disease characterized by somatic mutation of gene PIG-A at the level of stem hematopoietic cell. This process results in disorder of synthesis of glycosil phosphatidyl innozitol (GPI) anchor fixing numerous molecules on membrane of blood cells which protect blood cells from impact of complement. The international society of clinical cytometry (2010) proposed the guidelines of detection of clone of paroxysmal nocturnal hemoglobinuria among erythrocytes, granulocytes and monocytes. The original technique is proposed to evaluate the clone of paroxysmal nocturnal hemoglobinuria in reticulocyte population of blood using method of flow cytofluorometry. The sampling of 160 samples of blood of patients with clinical symptoms of paroxysmal nocturnal hemoglobinuria and anemia was analyzed. Two modes of gatedrawing were applied--using monoclonal antibodies to CD71 (receptor to transferrin) and reagent BD ReticCount. The high correlation was established between size of reticulocytic clone of paroxysmal nocturnal hemoglobinuria evaluated by CD71 and size of granulocytic and monocytic clone of paroxysmal nocturnal hemoglobinuria. The developed panel (CD71/CD235a/CD59) can be applied for screening and monitoring of paroxysmal nocturnal hemoglobinuria.

Urokinase stimulates production of matrix metalloproteinase-9 in fibroblasts with involvement of reactive oxygen species.

In cultured fibroblasts, urokinase stimulated expression of MMP-9 and generation of ROS, while antioxidant ebselen abolished the stimulating effect of urokinase on MMP-9 expression. sTNF-α produced similar and more pronounced stimulating effect. The data showed that urokinase could regulate MMP-9 expression via ROS generation in fibroblasts, which can play an important role in stimulation of their migration and development of constrictor (negative) vascular remodeling due to thickening of the adventitia.

Application of molecular modeling to urokinase inhibitors development.

Urokinase-type plasminogen activator (uPA) plays an important role in the regulation of diverse physiologic and pathologic processes. Experimental research has shown that elevated uPA expression is associated with cancer progression, metastasis, and shortened survival in patients, whereas suppression of proteolytic activity of uPA leads to evident decrease of metastasis. Therefore, uPA has been considered as a promising molecular target for development of anticancer drugs. The present study sets out to develop the new selective uPA inhibitors using computer-aided structural based drug design methods. Investigation involves the following stages: computer modeling of the protein active site, development and validation of computer molecular modeling methods: docking (SOL program), postprocessing (DISCORE program), direct generalized docking (FLM program), and the application of the quantum chemical calculations (MOPAC package), search of uPA inhibitors among molecules from databases of ready-made compounds to find new uPA inhibitors, and design of new chemical structures and their optimization and experimental examination. On the basis of known uPA inhibitors and modeling results, 18 new compounds have been designed, calculated using programs mentioned above, synthesized, and tested in vitro. Eight of them display inhibitory activity and two of them display activity about 10 µM.

Proteolytically inactive recombinant forms of urokinase suppress migration of endothelial cells.

Proteolytically inactive recombinant forms of urokinase (uPAHQ and amino-terminal fragment) inhibit spontaneous migration of endothelial cells; amino-terminal fragment also suppresses angiogenesis stimulated by basic fibroblast growth factor in vitro. These findings suggest the possibility of using synthesized proteolytically inactive recombinant forms of urokinase for the regulation of endothelial cell migration and suppression of neoangiogenesis.

[The standardization of diagnostic of paroxysmal night hemoglobinuria cytometry].

The flow cytometry becomes a more and more largely applied technique. However, the sufficient novelty of technique has no worked-out standards of diagnostic of many diseases. The lacking of external control of quality promotes development of large variety of approaches to diagnostic of diseases and impossibility to compare the study results from different laboratories. The paroxysmal night hemoglobinuria is an acquired clonal disease characterized by proliferation of stem cells with partial or total loss of expression of glykosylphosphosphatidyl inositol anchor needed to conjugate a number of surface proteins. The flow cytometry is a basic technique of detection and monitoring of clone of paroxysmal night hemoglobinuria. The article presents the results of paroxysmal night hemoglobinuria testing of 8 patients in 6 independent laboratories using flow cytometry by standard protocol recommended by the International society of clinical cytometrists (ICCS).

[Methods of detection and prognostic significance of genetic material increase of the short arm of chromosome 2 and a deletion of the short arm of chromosome 1 in patients with neuroblastoma].

MYCN gene amplification and 1p deletion in neuroblastoma patients are associated with poor prognosis and commonly used for patient's stratification into risk groups. MYCN copy number and 1p deletion status were analyzed with multiplex ligase-dependent probe amplification (MLPA), PCR and FISH. MYCN amplification was revealed in 21 patients (17.2%) simultaneously by MLPA and PCR. In 28 cases (23.0%) 2p gain was detected. 1p deletion was revealed in 28 patients (23.0%) while concordance between PCR and MLPA achieved 95.8%, PCR and FISH - 90.9%. Mean follow-up time achieved 42 months (ranged from 1 month to 13 years). Event-free survival and overall survival in MYCN-amplified patients as well as in patients with 1p deletion were significantly lower comparing with MYCN-negative patients or patients without 1p deletion.

[Regulation of growth and remodeling of blood vessels: the unique role of urokinase].

Urokinase-plasminogen activator (uPA) is a multifunctional fibrinolytic protein activating growth factors, inducing proteolytic cascades, modulating cytokines, regulating receptor shedding, cellular phenotypic modulation and protein expression. These mechanisms underlie the ability of uPA to stimulate the key processes of vascular remodelling, atherosclerosis progression, restenosis and angiogenesis, -- cell migration and proliferation. We summarized data received by us and others concerning the role of uPA in blood vessel remodelling and growth. At the present stage, the uPA may be considered as a perspective target for influences directed on both the prevention of negative arterial remodelling and restenosis as well as the stimulation of vessel growth at ischemic diseases.

Urokinase stimulates inflammatory response in damaged vascular wall during in vivo arterial remodeling.

Perivascular application of urokinase to the ballooned artery promoted the growth of neointima and constrictive remodeling of the vessel and stimulated the inflammatory response in the damaged vascular wall in vivo. Recombinant tissue plasminogen activator did not induce these changes. Our results indicate that urokinase is involved in the regulation of the inflammatory response during in vivo remodeling of the damaged vascular wall.

[Plasminogen activators and matrix metalloproteinases during arterial remodeling].

To evaluate the role and interaction of plasminogen activators and matrix metalloproteinases (MMPs) in arterial remodeling in vivo we compared effects of recombinant urokinase- (uPA) and tissue-type (tPA) plasminogen activators on vessel morphology, cell proliferation, inflammatory reaction and MMPs expression in arterial wall after experimental balloon angioplasty. We observed that the periadventitial application of uPA to the injured artery in pluronic gel stimulated neointima formation and inward arterial remodeling as well as cell proliferation and inflammatory leukocytes recruitment. In contrast, tPA attenuated neointima growth, contributed to outward arterial remodeling and did not affect significantly leukocytes recruitment in injured arterial wall. Perivascular uPA increased the content and activity of MMPs, while tPA did not induce such changes. In mouse model of vascular remodeling based on partial ligation of the carotid the content of uPA correlated with neointima growth, tPA content correlated with outward arterial remodeling. Our experiments suggest that plasminogen activators represent specific functional target for attenuating unfavorable inward arterial remodeling.

Urokinase induces ROS production in vascular smooth muscle cells.

Urokinase stimulates the production of superoxide radical in cultured aortal smooth muscle cells simultaneously with activation of the expression of NAD(F)H-oxidases nox1, nox4, and phox22. Antioxidant ebselen abolishes the stimulating effect of urokinase on smooth muscle cell proliferation. The data showed that urokinase can potentiate oxidative stress in the arterial wall and can play an important role in the development of adverse arterial remodeling.

Urokinase increases the content and activity of matrix metalloproteinases 2 and 9 during in vivo constrictive arterial remodeling.

Perivascular application of urokinase to ballooned artery stimulating the formation of neointima and constrictive arterial remodeling increased the content and activity of matrix metalloproteinases 2 and 9 in vivo. Application of recombinant tissue plasminogen produced no such changes. It was demonstrated that urokinase stimulates expression of matrix metalloproteinases in vivo.

[Expression of urokinase plasminogen activator, its receptor and plasminogen activator inhibitor type 1 in different types of atherosclerotic lesion in human aorta]. / Ekspressiia urokinazy, ee retseptora i ingibitora aktivatorov plazminogena 1-go tipa v stenke aorty cheloveka pri raznykh tipakh ateroskleroticheskogo porazheniia.

The role of plasminogen activators in the regulation of key processes of atherosclerosis progression stays unclear. The aim of this study was to evaluate the expression of urokinase plasminogen activator (uPA), its receptor (uPAR) and the plasminogen activator inhibitor type 1 (PAI-1) in human aorta, and to balance them with the stage of atherosclerotic lesion. We have shown that uPA and uPAR in normal aorta are mostly expressed by intimal smooth muscle cells. The expression of these proteins was up-regulated in diseased aorta compared to normal artery. The most part of cells in both fatty streak and fibro-fatty lesion were monocytes/macrophages, and about 60% of these cells expressed uPA and its receptor. PAI-1 was mostly localized on the lumonal part of the aorta and in the extracellular matrix of the intima. We observed a moderate increase of PAI-1 expression in atherosclerotic lesion. Thus, our data indicate participation of plasminogen system in atherogenesis.

Plasminogen activators in vascular remodeling and angiogenesis.

This review considers cellular and molecular mechanisms of the involvement of plasminogen activators in extracellular proteolysis and cell migration and proliferation. The role of plasminogen activators in vascular remodeling in atherosclerosis, restenosis, and angiogenesis is discussed.