[CIC-rearranged sarcoma: a case report and literature review]. / CIC-perestroennaya sarkoma: klinicheskoe nablyudenie i obzor literatury.

CIC-rearranged sarcoma is a rare and extremely aggressive tumor that occurs mainly in soft tissues. Despite the fact that identification of a characteristic genetic rearrangement is necessary to verify the diagnosis, in most cases, the correct diagnosis can be made by comparing histological signs and a characteristic immunophenotype, which greatly speeds up the diagnosis. The article describes a case of CIC-rearranged sarcoma in a 14-year girl with the successful application of the CWS-2009 treatment protocol.

Influence of Extracellular Vesicles from the Follicular Fluid of Young Women and Women of Advanced Maternal Age with Different miRNA Profiles on Sperm Functional Properties.

We studied the effect of co-culturing of extracellular vesicles in the follicular fluid of young women and women of advanced maternal age on sperm motility. Vesicles were obtained by differential centrifugation. The sperm fraction was isolated from the seminal fluid of 18 patients (age 28-36 years). The spermatozoa were incubated with vesicles (1:2 ratio) for 60 or 120 min at 37°C in a CO2 incubator. A fraction of spermatozoa incubated without vesicles served as the control. After the incubation, the sperm samples were sedimented by centrifugation, fixed in 2.5% glutaraldehyde, and analyzed by transmission electron microscopy. RNA was isolated from the follicular fluid vesicles by column method followed by cDNA synthesis in a reaction mixture according to miScript II RT Kit protocol (Qiagen). After 60-min incubation with extracellular vesicles from the follicular fluid of women of advanced maternal age, the sperm motility and hyperactivation slightly changed in comparison with the group where incubation was performed with follicular fluid vesicles from young women and control group. Follicular fluid miRNA profiles in women of different ages varied, which suggests different functional compositions and effects of follicular fluid vesicles of different age groups on sperm characteristics. Transmission electron microscopy revealed differences in the interaction of follicular fluid vesicles from women of different age groups with spermatozoa. Further study of the effect of extracellular vesicles from the follicular fluid and analysis of their transcriptomic, proteomic, and metabolomic composition on sperm mobility and fertilizing ability will improve the effectiveness of assisted reproductive technology programs in patients with male infertility.

Influence of Extracellular Vesicles of the Follicular Fluid on Morphofunctional Characteristics of Human Sperm.

We studied the effect of extracellular vesicles of the follicular fluid on morphofunctional characteristics of human spermatozoa using CASA (computer-assisted sperm analysis) analytical system. The vesicles were obtained by sequential centrifugation at different rotational speeds and frozen at -80°C in the Sydney IVF Gamete Buffer medium. The sperm fraction was isolated from the seminal fluid of 21 patients aged 27-36 years by differential centrifugation in a density gradient. The precipitate was suspended in Sydney IVF Gamete Buffer to a concentration of 106/ml and incubated with vesicles (1:2) at 37°C in a CO2 incubator for 30 min and 1 h. Sperm fraction incubated without vesicles served as the control. After incubation, some sperm samples were centrifuged at 700g for 5 min and fixed in 2.5% glutaraldehyde in 0.1 M buffer for transmission electron microscopy. After 30-min and 1-h incubation, the progressive and total sperm motility improved, the curvilinear and linear velocity of spermatozoa did not change significantly. Incubation with vesicles significantly changed the trajectory of sperm movement, which can attest to an increase in their hyperactivation and, probably, fertilizing capacity. Analysis of the effect of extracellular vesicles of follicular fluid on sperm motility will help to improve the effectiveness of assisted reproductive technology programs with male infertility factor by improving sperm characteristics in patients with asthenozoospermia and increasing the fertilizing ability of the sperm.

Laser Microtomography for IVF Oocyte of Human.

For in vitro fertilization technology, the quality of oocytes has a direct impact on the egg fertilization and developmental competence of early embryo. The morphological criteria are used for the estimation of oocyte quality before its fertilization in vitro. To date, only one method is known to determine the maturity of oocyte. This is the routine observation with a light microscope. The aim of this article was to adapt the noninvasive quantitative laser scanning microtomography (QLSM) for the investigation of morphological features of a human oocyte in vitro. This approach was used to accumulate the Z-stack gallery of optical sections of IVF oocyte. The layer-by-layer acquisition allows the fine cytoplasmic structure imaging. Applying the QLSM Z-stack of optical sections, the cellular volume was calculated with quantitative 3D reconstruction of a human oocyte. The volume value and intracellular structure were used as novel criteria to assess the oocyte state after the stress evoked by cryopreservation procedure.

Noninvasive Laser Microtomography of a Human Isolated Oocyte.

Using a non-invasive approach, quantitative laser scanning microtomography (QLSM), the morphology of human oocyte was studied layer-by-layer. Then, the cell volume was computed based on 3D reconstruction of a stack of optical sections obtained by QLSM. The integrity of oocyte membrane after cryopreservation was assessed by measuring the changes in oocyte volume in response to hypotonic shock.

Effect of Mechanical Microvibration on the Quality of Human Embryos during In Vitro Culturing and Outcomes of Assisted Reproduction Technologies.

We summarized the data on the role and efficiency of mechanical microvibration during embryo culturing in assisted reproduction protocols. The efficiency of culturing systems with microvibration in assisted reproduction programs and possible mechanisms of microvibration effect on the preimplantation development of human embryos are discussed.

Correction to: Comparative Analysis of the Expression of Glutathione Peroxidase and Glutathione Reductase Genes in Human Sperm after Cryopreservation.

The second author's name should read A. Yu. Romanov.

Comparative Analysis of the Expression of Glutathione Peroxidase and Glutathione Reductase Genes in Human Sperm after Cryopreservation.

We analyzed the effect of cryopreservation on the expression of glutathione peroxidase (GPX1) and glutathione reductase (GSR) genes in human sperm cells (15 sperm samples from fertile donors and 10 samples from infertile patients). The relative expression of GPX1 and GSR genes was determined by real-time PCR. The rate of post-thaw recovery was 2.1 times higher in the group of fertile donors. A significant increase in the expression of GPX1, but not GSR, was observed in sperm samples from infertile patients, while in patients with infertility, GPX1 expression significantly decreased after cryopreservation/thawing, in samples from fertile donors after the same procedure it increased to the level observed in the sperm samples from infertile patients. A positive correlation was revealed between GPX1 expression and sperm cryotolerance.

[Metabolomic profiling in culture media of day-5 human embryos]. / Profilirovanie metabolitov v pitatel'nykh sredakh piatidnevnykh émbrionov cheloveka.

The aim of this study was to determine the changes of metabolomic profiles in embryonic culture media (ECM) for the evaluation of quality and implantation potential of human embryos. ECM (n=163) were collected on day 5 before transfer or cryopreservation. Some embryos were used in preimplantation genetic screening for detection of aneuploidy karyotypes. Samples were subdivided into groups according to embryo morphological classification (by Gardner), genetic analysis and implantation data. ECM were extracted with methanol, precipitates were separated by centrifugation and metabolite production of individual embryo was analysed by LC-MS (the positive ion mode). After peak detection and retention time alignment, data were analysed using the PCA algorithm. MS fingerprinting analysis of embryo culture medium showed significant differences between morphologically divided groups. Intragroup comparisons did not reveal differences between subclasses. Genetic screening of embryos revealed 33 aneuploid karyotypes. It was shown that chromosome number did not affect the metabolite profiles comparing with the normal group. The culture media of embryos that were positive or negative for successful implantation showed specific signatures that allowed to distinguish embryos with different outcomes.The characterization of ECMs by LC-MS may facilitate more accurate selection of the best embryo for the implantation, improving single-embryo transfer and thus eliminating the risk and undesirable effects of multiple pregnancies.

[Comparative results of preimplantation genetic screening by array comparative genomic hybridization and new-generation sequencing].

Aneuploidies as quantitative chromosome abnormalities are a main cause of failed development of morphologically normal embryos, implantation failures, and early reproductive losses. Preimplantation genetic screening (PGS) allows a preselection of embryos with a normal karyotype, thus increasing the implantation rate and reducing the frequency of early pregnancy loss after IVF. Modern PGS technologies are based on a genome-wide analysis of the embryo. The first pilot study in Russia was performed to assess the possibility of using semiconductor new-generation sequencing (NGS) as a PGS method. NGS data were collected for 38 biopsied embryos and compared with the data from array comparative genomic hybridization (array-CGH). The concordance between the NGS and array-CGH data was 94.8%. Two samples showed the karyotype 47,XXY by array-CGH and a normal karyotype by NGS. The discrepancies may be explained by loss of efficiency of array-CGH amplicon labeling.

[Blastocyst Hatching in Humans].

The human oocyte is surrounded by the zona pellucida­an elastic, transparent extracellular matrix consisting of specific glycoproteins. The zona pellucida is preserved after fertilization and surrounds the developing human embryo for a few days. The embryo needs to get out of the zona pellucida before implantation to establish cell contacts between the trophectoderm and endometrial epithelium. The release of the embryo from the zona pellucida is carried out at the stage of the blastocyst and called zona hatching. During zona hatching the blastocyst breaks the zona pellucida and performs active movements to escape through a gap formed in the zona. While microscopic description of zone hatching is well known, biochemical and cytological basis of zone hatching remains poorly understood. The break of the zona pellucida occurs under the influence of two forces: mechanical pressure of the growing blastocyst on the zone and chemical dissolution of the zone material with secreted lytic enzymes. There is only one paper (Sathananthan et al., 2003), which describes the specialized cells in the trophectoderm that locally dissolve the zona pellucida, promoting the emergence of the hole for blastocyst release. Taking into account the singleness of the paper and the absence of further development of this subject by the authors in the following decade, the existence of specialized cells for zone hatching should be assumed with great care. Lytic enzymes, secreted by cells of the trophectoderm for dissolving the zona pellucida, are different. Depending on the species of the mammal, different classes of proteases participate in the zone hatching process: serine proteases, cysteine proteases, metalloproteinases. Proteases, secreted by human trophectoderm, are not described. The mechanisms of the active movement during blastocyst hatching are investigated to a lesser degree. Only the involvement of the cytoskeleton of trophectoderm cells in the mechanism of blastocyst compression was shown, and the participation of desmosomes in the coordinated change in the form of trophectoderm cells during compression is suggested. This review summarizes literature data on the possible mechanisms of zone hatching in the development of human embryos, obtained in experiments in vitro, as well as in animal models.

[Mechanisms of smooth endoplasmic reticulum aggregates creation in oocyte's cytoplasm in IVF cycles and its clinical relevance (literature review)].

A large proportion of human oocytes received from exogenous gonadotropin-stimulated cycles have different morphological attributes, or dysmorphisms. The presence of dysmorphism can affect the fertilization rate, the embryo quality and subsequently the frequency of occurrence of implantation and pregnancy. Special attention is paid to oocytes with cytoplasmic attributes such as alteration of cytoplasmic granularity, the appearance of vacuoles, lipofuscin bodies and visible (large) aggregates of smooth endoplasmic reticulum. Endoplasmic reticulum (ER) is a type of the organelle forming an interconnected network of flattened, membrane-enclosed sacs or tubes. One of the main functions of ER in the oocyte is storage and redistribution of calcium, which provides cell activation during fertilization. Furthermore, complex of ER and mitochondria is necessary for accumulation of energy, synthesis of lipids and triglycerides, as well as synthesis of cytosolic and nuclear membranes during the early stages of cleavage. The appearance of anomalously large aggregates of ER in oocytes correlates with a low fertilization rate, low embryo quality, and pregnancy rate. The aim of the manuscript is to summarize current understanding of the mechanism of formation of such pathology of oocytes, together with special aspects of their fertilization and embryo quality.

[Problem of pyoinflammatory diseases of soft tissues among military servicemen].

For a period of 2002-2009 according to indexes of morbidity was examined dynamics of morbidity of staff of Privolzhsko-Uralskij military region among soldiers and sergeants, of unit weight of PIDST, of unit weight of incapacity days, was observed seasonality. Was found a mid-year increase of index of morbidity among call-up staff on 6.84%, and among contract military servicemen--on 4.5%. PIDST take about 18% of incapacity days in structure of all workmisses of call-up military servicemen and about 7% among contract military servicemen. Was found a seasonality of one-wave type with a top of annual morbidity in August-October and decrease in first decade of November. There is a need of examination of immune status of persons of call-up age.

[Asymptomatic genital herpes infection and infertility in males].

Incidence of Herpes simplex virus (HSV) was studied in ejaculates of 100 men. The examinees had neither history nor clinical symptoms of HSV genital infection. HSV was detected by a rapid cultural method in the ejaculate of 20 out of 100 examinees (20%). Of 67 males with infertility HSV was detected in 25%, in 19 males examined prophylactically and 14 patients with varicocele (a comparison group) it was found in 10.5 and 7% cases, respectively. Oligozoospermia was two times more frequent in HSV-containing ejaculates than in HSV-negative one. Mean values of the majority of sperm parameters in HSV-positive and HSV-negative groups did not differ statistically. However, it was revealed that such damage of the spermatozoa structure as microhead (consequence of the defective acrosome or reduced genome) and cytoplasm drops on the neck (a sign of immature forms) occurred more often in HSV-infected patients than in persons with HSV-negative ejaculate. Thus, asymptomatic HSV infection has a negative effect on male fertility.

[Reparative regeneration of nerve fibers after sympathectomy: an experimental study].

The results of sympathectomies depend on the degree of regeneration of the sympathetic nerve trunk: the quicker the onset of regeneration the less remarkable the effect of operation. The aim of our experimental study WPS to examine the rate of regeneration of sympathetic nerve fibers softer different sympathectomy techniques (resection, electrocoagulation and cryodestruction of the part of the sympathetic nerve trunk) following 6 and 12 months. Nine random-bred rabbits were used (Central Research Laboratory, Urals State Medical Academy) to examine the rate of regeneration of the sympathetic nerve trunk contained by the vascular-nervous bundle of rabbit ear. Altogether 18 nerve sympathetic trunks were examined. The rate of regeneration was measured by morphometry. While examining the processes of reparative regeneration of the nerve fibers after resection, electrocoagulation and cryodestruction, account was taken of the changes in their quantity, diameter and degree of myelinization over time. The study of the morphologic picture during comparative analysis of regeneration made on different models of nerve derangement has demonstrated that after resection and electrocoagulation of the sympathetic trunk the process of regeneration proceeds most slowly. On cryogenic injury to the nervous tissue when only nerve fibers are damaged and connective tissue membranes preserve their anatomic integrity, there are created most favourable conditions for regeneration. Toward 12 months after cryodestruction the bundles of nerve fibers acquire an identical diameter, with predominance of the population of thick nerve fibers, and appear mature enough. After electrocoagulation the fibers contained by the bundles are different in diameter, with the predominance of medium-diameter fibers whereas the thick nerve fibers are not many. After resection the thick nerve fibers are lacking whereas the medium-diameter bundles contain myelinated nerve fibers, with the maintenance of the selective decay of the myelin membrane.

[Effects of sulfate mineral water and vetoron on the rate of radiation-induced aneuplodia in the bone marrow cells of rats]. / Vliianie pit'evoi sul'fatnoi mineral'noi vody i vetorona na chastotu vozniknoveniia radiatsionno-indutsirovannoi aneuploidii v kletkakh kostnogo mozga krys.

Experiments on non-inbred male rats have found that pretreatment with drinking sulfate mineral water and veteron reduced incidence of radiation-induced aneuplodia in bone marrow cells. The proportion of cells with normal number of chromosomes went up.

Does surgical correction of the superficial femoral vein valve change the course of varicose disease?

PURPOSE: We tested the hypothesis that the course of primary chronic venous insufficiency can be changed by correction of the incompetent valve in the superficial femoral vein. METHODS: This was a prospective, randomized, controlled study. A total of 125 extremities with reflux in greater saphenous and superficial femoral veins and belonging to clinical classes C2-C4 were analyzed. During the first 5 years, the type of clinical dynamics and changes in valvular function were determined in each patient. Patients were stratified according to the type of clinical dynamics and randomly assigned to treatment groups. Phlebectomy was performed in all extremities. In each of 64 extremities (the study group), the proximal incompetent superficial femoral vein valve was corrected as a part of the primary intervention. Patients were followed up for 7 to 8 years after surgery. RESULTS: The clinical dynamics of the extremity significantly depended on reflux changes. Increase in superficial femoral vein reflux was found in 74% of the extremities with progressive clinical dynamics but in only 47% of extremities with stable clinical dynamics (chi(2) = 9.71; P <.01). After surgical treatment, 65% of the extremities in the control group showed stable improvement; in 11%, recurrent varicosity was found; in 24%, the disease was aggravated. Of the extremities in the study group, 86% had stable improvement, 5% had recurrent varicosity, and 10% were aggravated (P <.05). Extremities with the progressive type of clinical dynamics were main contributors to this difference (chi(2) = 7.86; P <.05). In 92% of the extremities with corrected valves and in 50% of the extremities with increase in reflux after valvuloplasty, clinical improvement was observed (chi(2) = 11.5; P <.01). Extremities with corrected valvular function demonstrated superior results in comparison with extremities with stable valvular function in the control group (clinical improvement in 92% and 66% of extremities, respectively; P <.005). CONCLUSIONS: In patients with chronic venous insufficiency, two types of clinical dynamics, stable and progressive, can be identified. The progressive type is associated with the presence of superficial femoral vein reflux and increasing greater saphenous vein reflux. Superficial vein surgery neither corrects superficial femoral vein reflux nor prevents it from progressing further. Correction of a single superficial femoral vein valve during primary intervention significantly improves the long-term results of superficial venous surgery. This improvement is associated with the prevention of reflux progression. Surgical correction of the incompetent superficial femoral vein valve changes the course of primary chronic venous insufficiency.

[Selection of surgical tactics in colonic obstruction]. / Vybor khirurgicheskoi taktiki pri obturatsionnoi tolstokishechnoi neprokhodimosti.

From 1989 to 1996, 322 patients with colonic obstruction due to tumors underwent surgery in Surgical clinic No 1 of the Ural National Medical Academy. Specimens of 283 tumors removed at surgery (n = 213) or autopsy (n = 70) were examined in 322 patients. Complications were observed after 144 operations (44.7%); in 82 cases they were fatal (25.5% postoperative mortality). Postmortem examination was performed in 70 cases of 82 (85.37%). Histological examination of the 283 specimens of the resected prestenotic colon wall provided a histological classification of colonic wall insufficiency by identifying its compensation, subcompensation and decompensation stages. For possible prediction of a high risk for death in each specific case, a correlation analysis of 23 factors presumably influencing on outcomes was carried out. A decisive rule of prediction was formulated.

[Laparostomy in treatment of diffuse peritonitis]. / Laparostomiia v lechenii rasprostranennogo peritonita.

Two methods for laparostomy were used in peritonitis: programmed sanitation and variant of N.S. Makokha method (68.8 and 31.2% of patients respectively). 215 patients with diffuse peritonitis have been under control for the period from 1989 to 1997, beginning with the method of Makokha for laparostomy and then converting to programmed sanitation. Lethality rate in 1989-1992 made up 67.6%, in 1993-1997--35.4%. Among the patients, in whom for performing elective sanitation "ventrophiles" were fixed during the initial operation, lethality made up 25.7% and if it was done during the second and later procedures, lethality made up 45.7%.

[The prognosis of the immediate results of surgical treatment in colonic obstructive ileus of tumor origin]. / Prognozirovanie blizhaishikh rezul'tatov operativnogo lecheniia pri obturatsionnoi tolstokishechnoi neprokhodimosti opukholevogo geneza.

For making prognosis of high risk of the lethal outcome the authors used a correlative analysis of 23 factors influencing the treatment results. Ten of them have high correlation coefficients and reliably influence the level of postoperative lethality. The rule of the prognosing of the nearest results of operative treatment is formulated which allows the adequate assessment of the patient's state in dynamics and the timely correction of the process of treatment. The elaborated scheme of the score assessment is simple in use, informative and statistically reliable. It can be recommended for wide clinical practice.