Vol 23, No 3 (2020)

Gynecological morbidity particularly formation of menstrual function poses an important task because menstrual irregularities comprise a lead pathology among adolescent girls and considered as a marker of adverse environmental impact on female body [1, 2, 3]. The territories of the Orenburg Region [4] are known to be coupled to profound environmental and hygienic issues. We acknowledge the influence of anthropogenic stress on physical development and health in childhood by recording deviated physical development most frequently in children living in ecologically unfavorable territories [5]. Previously, we published the data on gynecological morbidity pattern in adolescent girls living in the Orenburg Region in territories with differed 7 year-monitored (from 2010 to 2016) anthropogenic stress [6]. Here we present the data on surveying girls aged 15-17 years with menstrual irregularities (MIs) living in the regions of the Orenburg region with varying anthropogenic stress assessed for their physical development, hormonal and immunological parameters. 2010- 2019 primary gynecological morbidity data were assessed based on available annual reports for various territories of the Orenburg Region. Subjects residing in the Western, Central and Eastern territories of the Orenburg Region were assesed based immune status data such as parameters of cell-mediated immunity, phagocytic indices, serum immunoglobulin class A, M, G as well as circulating immune complex levels. Hormone status was analyzed by assessing serum level of thyroid and sex hormones. It was found that oligomenorrhea in MIs was prevalent that tended to result in leptosomy and pachisomy in physical development. Most changes in adolescent girls were found in relative frequency of peripheral blood CD3⁺, CD4⁺ and CD19⁺ lymphocytes, phagocytic index as well as IgM, estradiol, progesterone, testosterone levels, LH, 17-OH-progesterone and cortisol. Our study demonstrated that adolescent girls living in the West and East of the region were featured with dominant oligomenorrhea in MIs associated with physical development tended to leptosomy and pachisomy. Assessing parameters of physical development in adolescent girls is of great importance and should be performed to diagnose potential menstrual irregularities. Thus, our data underline importance of negative impact of anthropogenic burden on gynecological health in adolescent girls.

Neonatal pneumonia is one of the most common causes of infant mortality. Moreover, the diagnosis of this pathology represents a difficult task and requires to seek for additional solutions in clinical and laboratory practice. One of the current and promising ways to diagnose rerlies on search of predictive markers among the innate and adaptive immunity genes involved in disease pathogenesis. We used bioinformatics analysis to select genes playing essential role developing neonatal pneumonia. Next, cord blood samples collected from 234 newborns were examined by real-time polymerase chain reaction for nine polymorphic markers in the TNF, IL10, IL17A, IL17F, IL6, NOS3, and MMP9 genes. The results of statistical analysis showed that heterozygous genotypes in the MMP9 and TNF genes were associated with the risk of developing early neonatal pneumonia, and also the protective role of homozygotes AA in the MMP9 gene and GG in the TNF gene. A search for associations with a risk of intrauterine pneumonia revealed an unfavorable role for heterozygous genotypes in the NOS3 and MMP9 genes. Thus, due to difficulties in diagnosis or in case of developing neonatal pneumonia, it may be recommended to add genetic analysis for assessing polymorphic markers in the MMP9 (rs17576), TNF (rs1800629) and NOS3 (rs1549758) genes along with standard test assays.

Cytokines are small-molecular weight proteins involved in regulating inflammation, proliferation and differentiation of immunocompetent cells, as well as effector functions. IL-1β and TNFα are among the most powerful inducers of the inflammatory response. Production of proinflammatory cytokines comprises normal response to infection and represents an arm of various autoimmune disease pathogenesis. It is worth comparing cytokine production in autoimmune and infectious diseases to determine features of cytokine profile. The aim of the study was to evaluate proinflammatory cytokines IL-1β and TNFα produced by blood cells in children with autoimmune and infectious diseases.
194 children, aged 2-17 years, were examined: 99 patients with juvenile idiopathic arthritis; 26 patients with unspecified reactive arthropathy; 14 children with systemic lupus erythematosus; 24 children with chronic viral hepatitis C; 33 healthy children. Heparinized blood samples were diluted with a glutamine-containing culture medium RPMI-1640, samples with/without phytohemagglutinin stimulation were prepared as well. Samples of diluted blood were incubated for 24 hours (37 °C, 5% CO₂). The concentrations of IL-1β and TNFα in the cell culture supernatants were determined by ELISA.
It was found that groups of patients with rheumatic diseases (systemic lupus erythematosus, juvenile idiopathic arthritis and unspecified reactive arthropathy) were featured with spontaneous production of IL-1β and TNFα at higher level than in control, and the stimulated synthesis of IL-1β was lower. In patients with chronic viral hepatitis C, the spontaneous concentration IL-1β and TNFα and the stimulated concentration of IL-1β did not differ those ones found in healthy children. Stimulated TNFα production in patients with juvenile idiopathic arthritis, unspecified reactive arthropathy, and hepatitis C was significantly higher than in control group. More intensive spontaneous production IL-1β and TNFα in groups of patients with rheumatic diseases indicates previous activation of immunocompetent cells. Decreased stimulated IL-1β production in groups with various diseases points at exhaustion of immunocompetent cell functional reserve due to chronic activation.

Interleukin 17 plays a key role in the immunopathogenesis of rheumatoid arthritis (RA) and serves as a link between activation of innate and adaptive immune cells, whereas its increased expression may represent one of the causes for uncontrolled inflammation and formation of immunopathological reactions. Among members of the interleukin 17 family, most studied is IL-17A, which is characterized by peak biological activity. IL-17A is one of the important immune mediators able to induce production of other pro-inflammatory cytokines and chemokines and promote recruitment of inflammatory cells, such as monocytes and neutrophils, into inflamed organs. IL17A gene contains a number of polymorphic sites, wherein single-nucleotide substitutions particularly at position the -197G/A may affect its expression level. Here in case-control study we retrospectively examined contribution of genetic polymorphism at the -197G/A position within the IL17A gene. Allele-specific PCR was used to iderntify the -197G/A polymorphism in IL17A gene in groups of patients with rheumatoid arthritis and healthy donors of the Russian ethnic group. Our study was made within a framework on assessing immunogenetic component for rheumatoid arthritis in ethnic Russian subjects in the Chelyabinsk Region. Prevalence of IL17A gene alleles and genotypes obtained in the work is in agreement with the Hardy– Weinberg equilibrium, and is characterized by rather high frequency of allele replacement (40%), which is typical for Caucasoid populations. Thus, it was found that interpopulation differences are characteristic of such gene polymorphism shown not to be associated with predisposition to rheumatoid arthritis in ethnic Russian subjects in the Chelyabinsk Region. Women with RA in our study were found to display certain changfes in frequencies of alleles and genotypes formed due to single-nucleotide substitution in IL17A gene at position -197G/A. However, in women such features cannot be considered as additional risk factors for developing RA. Allele -197*G, homozygous genotype -197G/G may be considered as markers of late-onset for the first RA attack in women. Analysis on distribution of SNP -197G/A alleles and genotypes within the IL17A gene showed that such polymorphism is of low value predictor likely being more associated with some RA clinical variants, but not with predisposition to RA development.

The gastrointestinal tract is a barrier to the penetration of allergens into the organism and can be one of the shock organs of the development of allergic inflammation. The diagnosis of gastrointestinal manifestations of allergies is not given due attention, since their clinical symptoms are not pathognomonic, but are identical to diseases of the gastrointestinal tract of various genesis. The aim: to study the features of the structural and functional state of the gastrointestinal tract and the spectrum of sensitization to food and inhalation allergens of children with gastrointestinal manifestations of atopy. The analysis of medical histories of children with gastrointestinal manifestations of allergies aged from 1 to 18 years (n = 28), who were treated at the Regional children's allergological center in Krasnoyarsk, was performed. All patients underwent a specific allergological examination (allergological history, determination of the concentration of total IgE, specific IgE to food and inhalation allergens). The assessment of the state of the gastrointestinal tract was carried out taking into account complaints, anamnesis, objective examination and the results of esophagogastroduodenoscopy. It is established that gastrointestinal manifestations of atopy were most often combined with dermatorespiratory manifestations of allergy in the form of allergic rhinitis, atopic bronchial asthma and atopic dermatitis. Among the lesions of the gastrointestinal tract with the highest frequency of occurrence, changes in the nature of the stool (diarrhea, constipation) and the presence of symptoms of inflammation of the gastrointestinal tract-gastroesophageal reflux, gastritis, bulbitis, duodenitis, were determined. According to the results of esophagogastroduodenoscopy, gastritis and cardia insufficiency were most often determined. According to the results of the analysis of the concentration of total IgE in the blood serum, the increased level was determined in 42,8% of cases, the average value of total IgE in the blood serum in patients with atopic genesis of allergies was 140,9 IU/ml. These data evidence that the immunopathological basis for triggering allergic inflammation of the gastrointestinal tract may be based not only on atopic mechanisms. Based on the analysis of specific IgE concentrations, the characteristics of the spectrum of sensitization to food and inhalant allergens of children with gastrointestinal manifestations of atopy were determined. The most significant allergens were: food – flour mix; pollen – mixed of trees; domestic – perennial mixed. Thus, when conducting a specific allergological examination of children with atopy, it is necessary to take into account the systemic nature of the process and pay special attention to assessing the state of the gastrointestinal tract, as one of the shock organs in the development of allergic inflammation.

This paper highlights the features of immune regulation and specific hypersensitivity in children residing in the Northern Siberia being exposed to the extreme climatic and exogenous chemical risk factors. An immunological diagnostic examination of 255 children permanently residing in the region of exogenous hapten exposure was conducted. The control group consisted of 130 children residing on the territory not being exposed to exogenous chemical factors. Level of serum class G, A, M immunoglobulins was assessed by using a Mancini radial immunodiffusion reaction. Amount of total serum IgE, neuron-specific enolase, and VEGF was measured by using the ELISA. Changes in level of IgG specific to phenol and copper as well as IgE specific to nickel and benz(a)pyrene were measured by using enzyme-linked allergosorbent test. Peripheral blood lymphocyte subsets were analyzed by using membrane-bound immunofluorescence with a panel of labeled specific monoclonal antibodies. High-quality statistical data analysis was performed by using Microsoft® Office Excel 2003 software as well as Statistica 6.0 application software package (StatSoft, USA). Statistical significance was determined by using Student’s t-test. Relation between signs was estimated by using correlation and regression analysis as well as Fisher’s test. Immunological study demonstrated that immunogram parameters were significantly changed in the observation group by containing increased percentage of CD16⁺ effector cells absolute and relative CD19⁺ cell count (p ≤ 0.05), relative and absolute count of CD25⁺ lymphocytes (p ≤ 0.05) as well as CD95⁺T cells as compared to the reference values and control group (p ≤ 0.05). Moreover, level of neuron-specific enolase and VEGF (vascular endothelial growth factor) was increased compared to control group (p ≤ 0.05). It points that regulatory mediators potentially mapped to the glia and vascular intima in children from the observation group were extremely activated. The level of both total (total IgE) and specific sensitization was significantly increased by assessing immunoglobulin G specific to phenol as well as immunoglobulin E specific to nickel and benz(a)pyrene (p < 0.05). Parameters of programmed cell death were significantly altered that was paralleled with simultaneous activation of general and specific sensitization highlighting features of the immunological status in children examined distinguished by excessive stress of immune regulatory arms in case of combined exposure to climate-geographic as well as exogenous chemical risk factors.

A close link between processes of immune-based and osteogenesis, influence of immune system on reparative processes after a traumatic femur fracture, as well as state of increased afferent signals emitted by intraosseous receptors and hyperstimulation of immune cells during trauma accounts for feasibility of using efferent multimodal therapy interventions, which may be accomplished by medical ozone. The aim of the study was to assess dynamic changes in pain syndrome and level of pro- and anti-inflammatory cytokines in isolated femur fracture treated with systemic ozone therapy.
The study group coprised 32 male patients with isolated femur fracture, average age 44.2±2.4 years. Starting from day 2 after surgical treatment, all patients received standard anticoagulant treatment and antibiotic therapy, 16 patients additionally received ozone therapy applied as small autohemotherapy (MAGT) at ozone therapeutic unit by using ozone destructor UOTA-60-01 “Medozon” manufactured by Medozon LLC, Moscow. Ozone concentration in ozone-oxygen mixture was 20 mg/l, per 10 ml-volume, applied as 7-9 injections course every other day. Patient-provided assessment of pain level in all groups was investigated by using a visual analogue pain scale. Concentrations of IL-6 and IL-4 cytokines were measured by using standard test systems (pruchased from Vector-Best JSC, Novosibirsk). Statistical data processing was carried out by using software package Statistica 10.0. The subjective pain level prior to the onset of the course ozone therapy and 2 days after surgical treatment was higher than average level after the end of MAGT course, pain level decreased to low level and significantly differed from that one before surgery, which indicates developed analgesic effect after systemic ozone therapy, likely associated with oxidative modification of inflammatory mediators. The level of pro-inflammatory cytokine IL-6 in patients with IPPK during systemic ozone therapy significantly decreased, which indicates some normalizing effect induced by ozone therapy on parameters of immunocyte secretory activity and prevents ovgert effects triggered by “cytokine storm”. Concentration of IL-4 vs after traumatic femur damage did not significantly differ among patient groups, except for differences with control group. Spearman correlation analysis revealed high degree of dependence between pain intensity and level of pro-inflammatory cytokines, whereas ozone therapy led to decreased pain intensity in lesion site and directly correlated with decreased IL-6 concentration. Thus, use of ozone at therapeutic concentrations limits excessive reactions of innate immunity that can lead to massive tissue damage in early stages of the posttraumatic process.

The current treatment strategy for rheumatoid arthritis has been formulated within the framework of the European League Against Rheumatism (EULAR) concept “Treatment to achieve the goal”. Methotrexate prescribed as soon as possible after verifying rheumatoid arthritis is recognized as a first-line drug that allows to achieve disease remission and prevent destructive changes in the joints. Long-term clinical experience of using methotrexate allowed to conclude that almost 30% patients with rheumatoid arthritis turn out to be resistant to such treatment, enforcing to change the basal anti-inflammatory therapy, shift to using targeted or genetically engineered biological drugs, so that timeframe to prevent disease progression can be irreversibly lost. In the last decade, genetic testing for drug therapy effectiveness has been gaining momentum based on individual features in functioning of enzyme systems which regulate various stages of drug biotransformation. To date, a personalized approach to treatment of rheumatoid arthritis may be implemented after examining more than a dozen of single nucleotide polymorphisms (SNPs) within the folate cycle genes responsible for metabolizing methotrexate as well as its mechanism of action. In our work, we attempted to test a relationship between therapeutic response (efficacy and resistance) to methotrexate and -401C>T (rs 3758149) SNP in the GGH (gamma-glutamyl hydrolase) gene, which coordinates the processes of extracellular methotrexate transport. A groups patients consisted of 85 basic anti-inflammatory therapy-naïve patients diagnosed with rheumatoid arthritis, who were initially treated with methotrexate at a dose of 10 to 17.5 mg per week, with subsequently assessed therapeutic efficacy 6 months after the treatment onset based on dynamics in DAS28 index that allowed to identify groups of “responders” and “non-responders”. Next, all patients from select groups underwent molecular genetic typing for GGH-401C>T SNP by using real-time polymerase chain reaction. Our study allowed to find that prevalence of the TT homozygous genotype (OR = 5.09; 95% CI 1.11- 23.3; p = 0.037) dominated in “methotrexate non-responders”, whereas “methotrexate responders” tended to have higher C allele frequency (OR = 0.54; 95% CI 0.27-1.01; p = 0.087), which allowed to identify them by genetic predictors of methotrexate therapeutic response in rheumatoid arthritis.

Patients with exacerbated chronic suppurative otitis media occurring in subjects with acute rhinosinusitis often consulted by ENT spacialists. A literature review has shown that much attention is paid to improving conservative therapy for such patients, and non-drug techniques remain applied rarely. In this context, based on a detailed study of biophysical characteristics, an opportunity of using low-frequency ultrasonic cavitation in combination therapy to increase local immunological resistance of mucous membranes in ENT organ cavities is justified. The aim of the study was to increase clinical therapeutic effectiveness of acute rhinosinusitis and exacerbated chronic suppurative otitis media based on examining timeframe necessary for normalization of pro-inflammatory cytokine profile after using low-frequency ultrasonic cavitation in combination therapy. 63 patients were enrolled to the study. The control group (n = 15) consisted of healthy volunteers, whereas main group had 48 patients with acute rhinosinusitis and exacerbated chronic suppurative otitis media, subdivided into 2 subgroups: subgroup 1 (n = 20) – patients receiving conventional treatment methods (elimination, unloading, local and systemic antibacterial therapy, puncture and anti-inflammatory treatment), subgroup 2 (n = 28) – patients underwent low-frequency ultrasonic cavitation (frequency 29 kHz, amplitude 25 μm) in combination therapy. Washouts from the surface mucous membranes of nasal cavity and nasopharynx were examined for level of IL-1β, IL-2, IL-8, IL-10, TNFα, IFNγ levels before treatment, at day 2, 7, 10 days and in one month after treatment.
Application of low-frequency ultrasonic cavitation in combination therapy in patients with acute rhinosinusitis associated with exacerbated chronic suppurative otitis media is accompanied by early recovery of balance in pro-inflammatory cytokines comparewd with conventional treatment methods. Data analysis showed that positive and satisfactory treatment result was achieved in 11 (56.2%) and 22 (78.2%) as well as 7 (36.2%) and 6 (21.8%) patients from subgroup 1 and 2, respectively. In two patients from subgroup 1 the result was noted as unsatisfactory.
Early recovery in balance of pro-inflammatory cytokines, improvement of clinical results and shortened treatment duration were observed after using low-frequency ultrasonic cavitation in combination therapy of patients with acute rhinosinusitis and exacerbated chronic purulent otitis. The data obtained may serve to justify introduction of low-frequency ultrasonic cavitation into combination conservative therapy of infected nasal and middle ear cavities as an effective and non-invasive approach.

Approximately 30 % of HIV-infected patients with viral load being suppressed during the course of antiretroviral therapy do not recover their peripheral CD4+T-lymphocyte counts. The reason for this phenomenon, named immunological non-response to treatment, remains unknown. In HIV-positive subjects receiving treatment, interleukin 7 plays a key role in increasing the number and supporting the viability of CD4⁺T-lymphocytes. We hypothesized that chronic immune activation, which develops in response to immunological failure during the therapy course, may reduce the susceptibility of CD4⁺T-cells to interleukin 7 in HIV-positive subjects. We examined 38 HIV-infected immunological non-responders to therapy; 42 HIVpositive patients with a standard response to treatment; 19 uninfected volunteers. The content of CD4⁺, CD4⁺CD127⁺ and activated HLA-DR⁺CD38⁺T-lymphocytes was determined in the peripheral blood of the examined individuals; the concentration of interleukin 7 was established. As a result, it was shown that interleukin 7 concentrations in the blood plasma of HIV positive immunological non-responders to treatment does not differ from the corresponding values of patients who gave a standard response to antiretroviral therapy. At the same time, immunological non-responders to treatment compared with subjects of both control groups were characterized by a deficiency of absolute and relative CD4⁺CD127⁺T-cell counts capable of responding to interleukin 7. Moreover, the interleukin 7 receptor expression level was reduced on CD4⁺T-lymphocytes of immunological non-responders. The higher was the frequency of activated CD4⁺T-lymphocytes; the lower was the CD127⁺ expression density. It should be noted that after excluding the data obtained from patients coinfected with HIV and hepatitis C virus, which are known to have significantly higher levels of chronic immune activation and systemic inflammation, we found no differences in CD127 expression between HIVpositive patients with distinct effectiveness of the immunological response to treatment. Thus, in the present study, we showed that in HIV-infection, poor immunologic response to antiretroviral therapy is associated with a decrease in the CD4⁺CD127⁺T-cell counts. Moreover, an increase in the level of chronic immune activation is associated with a decrease in CD127 expression density on CD4⁺T-lymphocytes.