Local Immune Status Research Articles

The influence of professional oral hygiene on the indicators of local immunity

Professional oral hygiene is an important part of measures aimed at preventing and maintaining the patient’s health. Local immune response is the first barrier of immune protection capable of preventing infection diseases. The aim of the study - the influence of professional hygiene on the indicators of local immunity has been studied. The study involved two groups of patients: the first group included 19 individuals with chronic caries of varying depths (class I), and the second group comprised 15 individuals with moderate periodontal disease. To evaluate nonspecific oral protection, the index of natural colonization of the buccal epithelium and the activity of microbial adsorption by oral mucosal cells were assessed. Specific immunity was evaluated by measuring the concentration of immunoglobulins in unstimulated saliva. All indicators were recorded both before and after professional oral hygiene procedures. When studying the index of natural colonization of the buccal epithelium, it was found that professional cleaning in both groups led to normalization of the spectrum of bacteria adhered to the epithelium: there was a decrease in the amount of gram-negative microflora, atypical for this biotope, while maintaining a high adhesion index of gram-positive cocci. In persons with caries, before sanation, “unsatisfactory” resistance of oral mucosa was revealed in a third of the examined, after sanation, the number of such patients decreased by 3 times, with “satisfactory” resistance increased by 2 times. In the group with periodontal disease, all patients exhibited “unsatisfactory” mucosal resistance before the hygiene procedures. After sanitation, 20 % “unsatisfactory” resistance persisted. In the majority of patients (80 %), the resistance of the oral mucosa improved to “satisfactory”. The concentration of all classes of immunoglobulins in both groups before sanitation was significantly below the norm, after the measures taken, the indicators increased significantly and reached the level typical for healthy individuals. Carrying out professional cleaning of the teeth leads to the normalization of local immune status even without additional medical immunocorrection.

DNA methylation variations of DNA damage response correlate survival and local immune status in melanomas.

We aimed to explore the impact of DNA methylation alterations on the DNA damage response (DDR) in melanoma prognosis and immunity. MATERIAL &METHODS: Different melanoma cohorts with molecular and clinical data were included. Hierarchical clustering utilizing different combinations of DDR-relevant CpGs yielded distinct melanoma subtypes, which were characteristic of different prognoses, transcriptional function profiles of DDR, and immunity and immunotherapy responsesbut were associated with similar tumor mutation burdens. We then constructed and validated a clinically applicable 4-CpG risk-score signature for predicting survival and immunotherapy response. Our study describes the close interrelationship amongDNA methylation, DDR machinery, local tumor immune status, melanoma prognosis, and immunotherapy response.

LOCAL IMMUNE STATUS OF PATIENTS AFTER HYSTEROSCOPIC MYOMECTOMY AND METHODS OF ITS CORRECTION

Aim. To evaluate changes in the local immune status of women after hysteroscopic myomectomy and the effect of the proposed treatment complex on them. Materials and methods. 221 patients with submucous uterine myoma who underwent hysteroscopic myomectomy were examined. 121 patients of the main group in the semi-operative period received complex rehabilitation therapy, 100 patients of the comparison group did not receive additional treatment after surgery. 80 women who applied for a routine preventive examination formed the control group. In all patients, the content of factors of local immunity – IL-6, IL-10, lactoferrin and secretory inhibitor of leukocyte proteinase in cervical mucus was studied by enzyme-linked immunosorbent assay (ELISA) using Atiscare reagents in the laboratory conditions of the Center. The indicators were studied before surgery, 3 and 6 months after surgery. Results. In patients with submucosal uterine myoma, an increased content of pro-inflammatory IL-6 in cervical mucus was found (357.9 pg/ml and 345.3 pg/ml), in the control group 134.7 pg/ml and a decrease in the concentration of anti-inflammatory IL-10 (37 .6 pg/ml and 34.3 pg/ml) while the value of the control group was 78.5 pg/ml. The increase in the cervical content of the secretory inhibitor of leukocyte proteinase in women with submucous uterine myoma (274.2 pg/ml and 284.9 pg/ml against 114.5 pg/ml in the control group) is evidence of an active reaction of the mucous membrane to the inflammatory process. The role of the factor in the implementation of antisperm immunity is a factor of impaired fertility in such women. Elevated lactoferrin (27.4 ng/ml and 28.4 ng/ml vs. 3.5 ng/ml in healthy women) is also associated with infertility. Performing hysteroscopic myomectomy leads to a gradual, but incomplete normalization of the content of the specified immune factors in the cervical mucus, and the improvement of the treatment complex leads to the full achievement of the indicator of the control group after 6 months, which is important for the implementation of reproductive function. Conclusions. 1. Patients with submucous uterine myoma are characterized by an increased content of pro-inflammatory factors of local immunity – IL-6, lactoferrin and secretory inhibitor of leukocyte proteinase, as well as a reduced concentration of anti-inflammatory IL-10. 2. After hysteroscopic myomectomy for 6 months, there is a decrease in the content of pro-inflammatory factors and an increase in the concentration of IL-10 in the cervical mucus, but the indicators do not reach the values of healthy women. 3. The use of the proposed complex of treatment after hysteroscopic myomectomy allows to achieve faster changes in the indicators of local cervical immunity, and after 6 months to achieve the values obtained in healthy women.

Burn Wound Healing Activity of Hydroxyethylcellulose Gels with Different Water Extracts Obtained from Various Medicinal Plants in Pseudomonas aeruginosa-Infected Rabbits.

Burn injuries represent a significant problem in clinical practice due to the high risk of infection and the prolonged healing process. Recently, more attention has been given to natural remedies such as water extracts of various medicinal plants, which possess anti-inflammatory and wound healing properties. The aim of this study is to evaluate the efficacy and safety of Satureja montana L. and other water extracts in a burn wound model. The study involved male Californian rabbits (n = 52) divided into eight groups. Burn wounds were modeled on the animals and subsequently treated with gels based on Satureja montana L. and other water extracts. The reparative potential of the epidermis (assessed by Ki-67 expression), the state of local immunity (measured by the number of CD-45 cells), and the anti-inflammatory role of mast cells (measured by tryptase levels) were evaluated. Bacteriological and morphological studies were conducted. The most pronounced bactericidal, reparative, and immunostimulatory effects were observed after the treatment using a gel mixture of water extracts from Satureja montana L., Salvia sclarea, Coriandrum sativum L., and Lavandula angustifolia in equal proportions (1:1:1:1). The other gels also demonstrated high efficacy in treating burn wounds, especially when using a strain of Pseudomonas aeruginosa resistant to several antibiotics. Immunohistochemical studies showed a significant increase in the number of Ki-67-positive cells in the basal layer of the epidermis and a decrease in the number of CD-45-positive cells, indicating improved proliferative activity and reduced inflammation. This study confirms the hypothesis that the use of water extract mixtures significantly enhances the reparative potential, improves the immune response in the treatment of burns, and promotes wound healing. These findings pave the way for further research and the application of complex phytotherapeutic agents, specifically water extracts of medicinal plants containing phenols and antioxidants in burn wound therapy.

Revealing the novel metabolism-related genes in the ossification of the ligamentum flavum based on whole transcriptomic data.

The ossification of the ligamentum flavum (OLF) is one of the major causes of thoracic myelopathy. Previous studies indicated there might be a potential link between metabolic disorder and pathogenesis of OLF. The aim of this study was to determine the potential role of metabolic disorder in the pathogenesis of OLF using the strict bioinformatic workflow for metabolism-related genes and experimental validation. A series of bioinformatic approaches based on metabolism-related genes were conducted to compare the metabolism score between OLF tissues and normal ligamentum flavum (LF) tissues using the single sample gene set enrichment analysis. The OLF-related and metabolism-related differentially expressed genes (OMDEGs) were screened out, and the biological functions of OMDEGs were explored, including the Gene Ontology enrichment analysis, Kyoto Encyclopedia of Genes and Genomes enrichment analysis, and protein-protein interaction. The competing endogenous RNA (ceRNA) network based on pairs of miRNA-hub OMDEGs was constructed. The correlation analysis was conducted to explore the potential relationship between metabolic disorder and immunity abnormality in OLF. In the end, the cell experiments were performed to validate the roles of GBE1 and TNF-α in the osteogenic differentiation of LF cells. There was a significant difference of metabolism score between OLF tissues and normal LF tissues. Forty-nine OMDEGs were screened out and their biological functions were determined. The ceRNA network containing three hub OMDEGs and five differentially expressed miRNAs (DEmiRNAs) was built. The correlation analysis between hub OMDEGs and OLF-related infiltrating immune cells indicated that metabolic disorder might contribute to the OLF via altering the local immune status of LF tissues. The cell experiments determined the important roles of GBE1 expression and TNF-α in the osteogenic differentiation of LF cells. This research, for the first time, preliminarily illustrated the vital role of metabolic disorder in the pathogenesis of OLF using strict bioinformatic algorithms and experimental validation for metabolism-related genes, which could provide new insights for investigating disease mechanism and screening effective therapeutic targets of OLF in the future.

Heyndrickxia coagulans strain SANK70258 suppresses symptoms of upper respiratory tract infection via immune modulation: a randomized, double-blind, placebo-controlled, parallel-group, comparative study.

Probiotic consumption strongly influences local intestinal immunity and systemic immune status. Heyndrickxia coagulans strain SANK70258 (HC) is a spore-forming lactic acid bacterium that has immunostimulatory properties on peripheral tissues. However, few reports have examined the detailed effectiveness of HC on human immune function and its mechanism of action. Therefore, we conducted a randomized, double-blind, placebo-controlled, parallel-group study to comprehensively evaluate the effects of HC on immunostimulatory capacity, upper respiratory tract infection (URTI) symptoms, and changes in intestinal organic-acid composition. Results of a questionnaire survey of URTI symptoms showed that runny nose, nasal congestion, sneezing, and sore throat scores as well as the cumulative number of days of these symptoms were significantly lower in the HC group than in the placebo group during the study period. Furthermore, the salivary secretory immunoglobulin A (sIgA) concentration was significantly higher, and the natural killer (NK) cell activity tended to be higher in the HC group than in the placebo group. In addition, we performed an exposure culture assay of inactivated influenza virus on peripheral blood mononuclear cells (PBMCs) isolated from the blood of participants in the HC and placebo groups. Gene-expression analysis in PBMCs after culture completion showed that IFNα and TLR7 expression levels were significantly higher in the HC group than in the placebo group. In addition, the expression levels of CD304 tended to be higher in the HC group than in the placebo group. On the other hand, the HC group showed a significantly higher increase in the intestinal butyrate concentration than the placebo group. HC intake also significantly suppressed levels of IL-6 and TNFα produced by PBMCs after exposure to inactivated influenza virus. Collectively, these results suggest that HC activated plasmacytoid dendritic cells expressing TLR7 and CD304 and strongly induced IFNα production, subsequently activating NK cells and increasing sIgA levels, and induced anti-inflammatory effects via increased intestinal butyrate levels. These changes may contribute to the acquisition of host resistance to viral infection and URTI prevention.

Dickkopf-related protein 1 as a biomarker of local immune status and worse prognosis of Oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is an infiltrative malignancy characterized by a significantly elevated recurrence rate. Dickkopf-related protein 1 (DKK1), which plays an oncogene role in many cancers, acts as an inhibitor of the Wingless protein (Wnt) signaling pathway. Currently, there is a lack of consensus regarding the role of DKK1 in OSCC or its clinical significance. To examine the role and effect of DKK1 in OSCC. The identification of differentially expressed genes (DEGs) in OSCC was conducted by utilizing databases such as The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO). A comprehensive analysis of gene expression profile interactions (GEPIA) and Kaplan-Meier curve were conducted to investigate the associations among DEGs, patient survival and prognosis in individuals with OSCC. The biological function of DKK1 in OSCC was investigated by using molecular biology approaches. The expression of DKK1 was found to be upregulated in OSCC tissues at various stages. High levels of DKK1 expression exhibited a positive correlation with the overall survival (OS) and progression-free survival (PFS) rates among OSCC patients. DKK1 knockdown suppressed the proliferation and induced apoptotic response in OSCC cells. Moreover, DKK1 exerted a positive regulatory effect on HMGA2 expression, thereby modulating cell growth and apoptosis in OSCC. The expression of DKK1 was found to be positively correlated with the infiltration of immune cells in patients with OSCC. Additionally, higher levels of CD4 + T cells were associated with improved 5-year survival rates. DKK1 is a prognostic biomarker for patients with OSCC.

Стан місцевого імунітету в породілей з інфекцією акушерської хірургічної рани в умовах війни

Purpose - to study indicators of local immunity in the wound of women giving birth with an infection of an obstetric surgical wound in the conditions of war, which will give an opportunity to improve the therapy of wound infection at the present time. Materials and methods. Local immunity in the wound of 66 parturients with infected wounds during the war (the Group I) was studied and compared with indicators of local immunity in the wound of 76 parturients with infected wounds in peacetime (the Group II) six years to 2022. In addition, 15 healthy parturients - clinical remission (the Group III) were examined to determine the features of the course of the wound process. To study local immunity, the number of immunocompetent cells and phagocytic activity in biopsies from wounds, fixed immunoglobulins in smears-imprints were determined using the direct immunofluorescence method. Statistical data processing was carried out with the use of the application program package "Microsoft Excel 5.0" and the standard version "Statistica 8.0". The difference in indicators at Р<0.05 was considered statistically probable. Results and conclusions. In the examined women in labor with an infection of an obstetric surgical wound in the conditions of war under stressful conditions, there is a greater suppression of local immunity than in women in labor with this pathology in peacetime, which is expressed in the insufficient migration of immunocompetent cells to the focus of inflammation of the wound, especially B-lymphocytes, in the predominance of smears-imprints of plasma cells producing immunoglobulins of classes M and G, which lead to increased inflammation, inhibition of reparative processes in the wound, progression of the purulent-inflammatory process with the risk of septic complications during martial law, which leads to the correction of purulent wound therapy in obstetric practice during the war . The research was carried out in accordance with the principles of the Declaration of Helsinki. The research protocol was approved by the Local Ethics Committee of the institution mentioned in the work. Informed consent of the patients was obtained for the research.

Local periodontal immune status of patients after new coronavirus infection

The aim of the study was to identify indicators of cytokine and free radical periodontal status of patients with moderate severity of periodontitis that occurred after a new coronavirus infection 1–3 monthes ago. IL‑1ß, IL‑6, IL‑10, IL‑17A, nitrate-nitrite concentration, antioxidant activity, matrix metalloproteinase MMP‑8 in gingival fluid samples were studied. A significant increase in the local level of IL‑6, matrix methylproteinase‑8 and nitrate-nitrites was found in patients with periodontitis after a new coronavirus infection compared with the indicators of patients with periodontal diseases of the same severity, who have not had covid (p < 0.05). Patients with periodontitis after undergoing new coronavirus infection to develop comprehensive methods of therapy and prevention of complications.

Peculiarities of local immunity in dry eye disease on the background of hormonal dysfunction.

Aim: to determine the state of local immunity in DED on the background of hormonal dysfunction. Materials and Methods: Of 32 women, 17 patients with diagnosed SM and 15 women of the control group were examined. The Ocular Surface Disease Index and the state of local immunity were defined by determining Ig As in lacrimal fluid (LF) by radial immunodiffusion in Mancini agar. Results: During the OSDI questionnaire, a mild degree of DED was detected in 21 (65.6%) women, and an average degree was observed in 11 (34.4%) patients with SM. On average, OSDI was 34.54 ±2.01. As a result of studies of the state of local immunity in patients with SM, a tendency to increase Ig As was noted, compared with the control group. An increase in Ig As in the lacrimal fluid in patients with SM to 0.34 ±0.09 g/l was found, compared with the control group (0.24 ±0.03 g/l). Conclusions: Using the OSDI questionnaire, the presence of DED was detected in women with SM, mainly mild and moderate degree. The obtained results of the state of local immunity indicate in favor of a nonspecific inflammatory process, accompanied by a decrease in local immune protection and leading to further changes in the ocular surface.

Disorder of the Microbiota and Mucosal Immunity of the Genital Tract in Women of Reproductive Age with Different Clinical Courses of Acute Uncomplicated Pyelonephritis

The research deals with the study of immuno-microbiological parallels of the genital tract in women with acute uncomplicated pyelonephritis with concomitant chronic pelvic inflammatory diseases. In addition to etiotropic antibacterial therapy, correction of the mucosal immunity state of the genital tract is crucial for the treatment of the disease and prevention of its recurrence. The purpose of the work was to investigate the contamination and state of local immunity of the genital tract in women with acute uncomplicated pyelonephritis with different clinical courses. Methods. 246 women of reproductive age suffering from acute uncomplicated pyelonephritis were studied, divided into three variants of the disease clinical course: group 1 - mild (n=105), group 2 - moderate (n=90), and group 3 - severe (n=51) courses. The scrapings of the mucous membrane of the cervical canal and washings from the vagina were obtained before the start of antimicrobial therapy. Quantitative determination of pathogens was carried out by sowing the biological material on solid nutrient environment. Identification of detached bacteria was carried out according to Bergeys. Mollicutes (Ureaplasma spp., Mycoplasma hominis) were determined by the cultural-fermentative method using test systems and the polymerase chain reaction. Vaginal washings of 121 sick women were used for immunological studies. The levels of myeloperoxidase, lysozyme, human β-defensin-2, immunoglobulins M, A, G, secretory IgA, lactoferrin, C3-component of complement, secretory component, and tumor necrosis factor-α were determined. The reference group consisted of 23 clinically healthy women. The Statistica 12.0 program package for Windows was used, and a difference of p<0.05 was considered verifi able. Results. In the majority of patients with acute uncomplicated pyelonephritis (85.6% of cases), concomitant chronic pelvic inflammatory diseases (colpitis, salpingitis, underlying medical condition of cervix) were observed. Classical bacteria were more often detected in vaginal washings of patients of all groups, while mollicutes were detected in scrapings of the mucous membrane of the cervical canal, whereas the frequency of their detection increased with increase in the pyelonephritis severity. The analysis of indicators of local immunity determined disorders of mucosal immunity of the genital tract in the form of increased levels of myeloperoxidase, IgA, and IgG relative to reference values. The highest levels of lysozyme and myeloperoxidase were determined in patients with a severe course of acute uncomplicated pyelonephritis (by 5.3 and 3.6 times more, respectively), and the content of immunoglobulins A, M, and G in patients with a mild course of the disease (by 3.2, 3.1, and 4.0 times more, respectively). An increase in the level of tumor necrosis factor-α was detected in all patients, although no significant differences from controls were found in any group, but the highest median value was recorded in patients with severe pyelonephritis. In the group of patients with a severe clinical course, the lowest median was observed for β-defensin-2, which makes it impossible to inhibit the synthesis of TNF-α and, thus, supports the inflammatory process. Conclusions. It has been proven that in women with acute uncomplicated pyelonephritis, the severity of the course probably correlates with infection of the genital tract by mollicutes, mainly in association with classical bacteria and with level of mucosal immunity disorder. The obtained results of microbiological and immunological studies of biological material collected before the start of antibacterial therapy in patients with acute uncomplicated pyelonephritis proved the need for a mandatory examination by a gynecologist to ascertain concomitant chronic pelvic inflammatory diseases and establish infection with mollicutes to provide relevant etiotropic treatment. The above is the basis for adding the data regarding laboratory examination and treatment to the management protocols of patients with acute uncomplicated pyelonephritis.

Differences in the skin microbiota spectrum and parameters of local immunity in the areas of inflammation in skin diseases and healthy people

Alteration of microbiota composition is a trigger, and, sometimes, an etiological factor in the development of chronic skin diseases, e.g., psoriasis or eczema. Recognition of microflora by keratinocytes and immune cells leads to the production of antimicrobial peptides, chemokines and growth factors, which contribute to the differentiation of T lymphocytes to autoaggressive effector cells of Th1, Th17 and Th22 types that implement autoimmune inflammation in the psoriatic plaque. The aim of our work was to study the differences in the skin microbiota spectrum and the parameters of local immunity in capillary blood taken near the focus of inflammation in patients with autoimmune (psoriasis) and allergic (eczema) diseases compared with the parameters of healthy people. 24 patients with psoriasis (group 1), 20 patients with eczema (group 2) and 20 healthy adults (group 3) were examined. Biological materials were taken, i.e., the smears taken with sterile dry swab to the Amies transport medium with activated carbon, and capillary blood was taken in 2 microvets, 200 µL each) from the foci of inflammation on affected skin from the hands of patients, or from the fingers of healthy people. Inoculations of diagnostic media, microscopy with Gram staining and microbial identification were performed in a microbiological analyzer. Immunophenotyping of 22 subsets of mononuclear cells was performed by four-color staining of capillary blood with erythrocyte lysis and evaluation of subsets by a flow cytometer. Cytokines in blood plasma were determined by multiplex method. The spectrum of hand skin microflora of the group 3 was more diverse in the species composition. In patients with psoriasis and eczema, the coccal flora dominated, with a shift towards pathobionts in the microbiota spectrum. Activation of T and B cells, production of pro-inflammatory cytokines, IL-23/IL-17/IL-22 axis cytokines and cytokines – markers of epithelial cell damage (IL-25 and IL-33), as well as anti-inflammatory factors insufficiency were detected. Differences in changing parameters of the local immune status in patients with autoimmune (psoriasis) and allergic (eczema) diseases were revealed, thus reflecting the distinct features of immunopathogenesis in these diseases.

Construction of a Matrix Cancer-Associated Fibroblast Signature Gene-Based Risk Prognostic Signature for Directing Immunotherapy in Patients with Breast Cancer Using Single-Cell Analysis and Machine Learning.

Cancer-associated fibroblasts (CAFs) are heterogeneous constituents of the tumor microenvironment involved in the tumorigenesis, progression, and therapeutic responses of tumors. This study identified four distinct CAF subtypes of breast cancer (BRCA) using single-cell RNA sequencing (RNA-seq) data. Of these, matrix CAFs (mCAFs) were significantly associated with tumor matrix remodeling and strongly correlated with the transforming growth factor (TGF)-β signaling pathway. Consensus clustering of The Cancer Genome Atlas (TCGA) BRCA dataset using mCAF single-cell characteristic gene signatures segregated samples into high-fibrotic and low-fibrotic groups. Patients in the high-fibrotic group exhibited a significantly poor prognosis. A weighted gene co-expression network analysis and univariate Cox analysis of bulk RNA-seq data revealed 17 differential genes with prognostic values. The mCAF risk prognosis signature (mRPS) was developed using 10 machine learning algorithms. The clinical outcome predictive accuracy of the mRPS was higher than that of the conventional TNM staging system. mRPS was correlated with the infiltration level of anti-tumor effector immune cells. Based on consensus prognostic genes, BRCA samples were classified into the following two subtypes using six machine learning algorithms (accuracy > 90%): interferon (IFN)-γ-dominant (immune C2) and TGF-β-dominant (immune C6) subtypes. Patients with mRPS downregulation were associated with improved prognosis, suggesting that they can potentially benefit from immunotherapy. Thus, the mRPS model can stably predict BRCA prognosis, reflect the local immune status of the tumor, and aid clinical decisions on tumor immunotherapy.

CATION COMPOSITION OF ORAL SECRETION IN PRIMARY SCHOOL-AGED CHILDREN

SUMMARY. Recurrent respiratory infections (RRIs) currently have the greatest medical and social burden among infectious diseases worldwide, with the highest prevalence and incidence rates of acute respiratory infections in children. Recent studies indicate the role of inadequate immune response and the development of recurrent acute respiratory infections (ARI) in children due to deficiencies in essential trace elements in the body, which are crucial for the proper functioning of physical barriers and the immune system. The hypothesis of our study assumed that, considering the recurrence of at least 6 episodes of ARI per year, children with RRIs may have changes in salivary ion levels, which could serve as a basis for local immune suppression of the upper respiratory mucosa. The aim – to investigate the levels of metals in the saliva of children of primary school age with recurrent respiratory infections in comparison with episodically ill peers and establish the relationship of these changes with the state of local immunity. Material and Methods. In the actual study, 40 children participated, with 30 of them (the main group) being children with recurrent respiratory infections and 10 other children who had episodic illnesses were assigned to the control group. The levels of metal cations in the saliva were examined using inductively coupled plasma atomic emission spectrometry, including essential metals such as copper, manganese, zinc, calcium, cobalt, potassium, magnesium, sodium, phosphorus, selenium, iron, chromium, and sulfur. Additionally, levels of conditionally essential metals like lithium, nickel, and boron were measured. The levels of secretory IgA and lysozyme in saliva were determined using the enzyme-linked immunosorbent assay method. Results. The main group was not homogeneous; therefore, we divided it into two clusters. Respondents in one cluster had significantly lower relative levels of essential ion content in saliva. Specifically, calcium was lower in 64 % (p=0.003), copper in 76.5 % (p=0.001), manganese in 41.1 % (p=0.008), magnesium in 64 % (p=0.02), and zinc in 58.8 % (p=0.027). This fact can be explained by their increased losses due to frequent inflammatory processes in the upper respiratory tract, resulting in higher daily replenishment needs compared to their peers. Correlation analysis revealed a connection between illness incidence and the levels of copper (p=0.008), sodium (p=0.00005), and phosphorus (p=0.028), indicating potential associations between saliva metal profiles and the frequency of ARI in children with RRIs. The presence of dental caries correlated with lithium (p=0.02), copper (p=0.032), and nickel levels (p=0.037). Integral indicators of local immunity, such as lysozyme and secretory IgA, exhibited correlations with metal levels in saliva (Li, Cu, Ni, Na, Co, P, Zn), confirming the hypothesis of their role in regulating immune reactions in the oral cavity. Conclusions. Cluster analysis of the metal profiles of the group of children with RRI shows its heterogeneity, 43 % of representatives of this cohort had significantly lower levels of calcium (p=0.003), copper (p=0.001), manganese (p=0.008), magnesium (p=0.02) and zinc (p=0.027) than the control group. Which may indicate the presence of deficient conditions in some children with PRI as a result of the increased need for their recovery. The presence of caries, the level of SIgA, lysozyme (clinical indicators of local immunity of the oral cavity) have certain relationships with the levels of essential and conditionally essential levels of salivary metals (Li, Cu, Ni, Na, Co, P, Zn), which indicates the benefit of the connection between the metal profile of saliva and the state of local immunity.

Content of mediators of innate immunity in the tears of patients with vascular and neurodegenerative manifestations of diabetic retinopathy

According to the results of recent studies, diabetic retinopathy can be considered not only as a vascular disease, but also as a neurodegenerative process. Study of the composition of the tear fluid is used to assess the state of local immunity in the development of eye diseases. However, studies examining the effect of tear composition in diabetic retinopathy are few. The aim of the study is to determine the levels of IL-1β, IL-10, TGF-β3, MMP-7, TIMP-2, protein S100b, BDNF and NGF in the tear fluid ofpatients with vascular and neurodegenerative manifestations of diabetic retinopathy. The study included 80 patients diagnosed with type 2 diabetes which were divided into 2 groups: the 1st group included 40 patients who had no clinical signs of diabetic retinopathy on the fundus; the 2nd group included 40 patients with initial signs of non-proliferative diabetic retinopathy. All those included in the study were examined on an optical coherent tomograph RTVue-100 (USA); the volume of focal losses of retinal ganglion cells (FLV) was determined. An increase in FLV above the normative base of the device was regarded as an OCT-sign of retinal neurodegeneration. According to the results of OCT, the participants of the first and second groups were additionally divided into 4 subgroups: 1A — patients without vascular changes in the fundus and without OCT signs of retinal neurodegeneration (n = 12); 1B — patients without vascular changes in the fundus and with the presence of OCT signs of retinal neurodegeneration (n = 28); 2A — patients with initial non-proliferative DR and without OCT signs of retinal neurodegeneration (n = 10); and 2B — patients with initial non-proliferative DR and with OCT signs of retinal neurodegeneration (n = 30). The levels of IL-1β, IL-10, TGF-β3, MMP-7, TIMP-2, protein S100 b, BDNF, and NGF in tear fluid were determined by enzyme-linked immunosorbent assay. Levels of IL-1β and IL-10 in tear fluid in all subgroups were comparable to controls throughout the study. TGF-β3 content in the tear fluid of patients in the group with initial signs of non-proliferative DR (group 2) was significantly (p = 0.001) lower compared with control and group 2. However, there was no significant difference (p > 0.05) between subgroups A and B within groups. The concentration of MMP-7 in the tear fluid in all subgroups was significantly lower than in the control (p < 0.05). However, in the subgroups with OCT signs of retinal neurodegeneration (1B and 2B), the deficiency of this metalloproteinase was more pronounced (p = 0.0001). The levels of the neuropeptides under study NGF, BDNF and S100 B in tear fluid did not differ from controls in all subgroups.

Establishment and verification of prognostic model and ceRNA network analysis for colorectal cancer liver metastasis

ObjectsColorectal cancer (CRC) is one of the most common cancers in the world. Approximately two-thirds of patients with CRC will develop colorectal cancer liver metastases (CRLM) at some point in time. In this study, we aimed to construct a prognostic model of CRLM and its competing endogenous RNA (ceRNA) network.MethodsRNA-seq of CRC, CRLM and normal samples were obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus database. Limma was used to obtain differential expression genes (DEGs) between CRLM and CRC from sequencing data and GSE22834, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes functional enrichment analyses were performed, respectively. Univariate Cox regression analysis and lasso Cox regression models were performed to screen prognostic gene features and construct prognostic models. Functional enrichment, estimation of stromal and immune cells in malignant tumor tissues using expression data (ESTIMATE) algorithm, single-sample gene set enrichment analysis, and ceRNA network construction were applied to explore potential mechanisms.ResultsAn 8-gene prognostic model was constructed by screening 112 DEGs from TCGA and GSE22834. CRC patients in the TCGA and GSE29621 cohorts were stratified into either a high-risk group or a low-risk group. Patients with CRC in the high-risk group had a significantly poorer prognosis compared to in the low-risk group. The risk score was identified as an independent predictor of prognosis. Functional analysis revealed that the risk score was closly correlated with various immune cells and immune-related signaling pathways. And a prognostic gene-associated ceRNA network was constructed that obtained 3 prognosis gene, 14 microRNAs (miRNAs) and 7 long noncoding RNAs (lncRNAs).ConclusionsIn conclusion, a prognostic model for CRLM identification was proposed, which could independently identify high-risk patients with low survival, suggesting a relationship between local immune status and prognosis of CRLM. Moreover, the key prognostic genes-related ceRNA network were established for the CRC investigation. Based on the differentially expressed genes between CRLM and CRC, the prognosis model of CRC patients was constructed.

Interleukin-35 expression promotes hepatocellular carcinogenesis by inducing γδ T-cell exhaustion

Interleukin (IL)-35, a new member of the IL-12 family, exerts immunosuppressive effects in the hepatic microenvironment. Innate immune cells, such as γδ T cells, have vital roles in hepatic diseases, including acute and chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). In the current study, we focused on the effects and mechanisms of IL-35 on the local immune status of γδ T cells, especially in liver tumors. Based on CCK8 assay and immunofluorescence results, we showed that exogenous IL-35 stimulation of γδ T cells attenuated proliferative ability and killing functions against Hepa1–6 or H22 cells. Flow cytometry results showed that exogenous IL-35 stimulated the expression of programmed cell death 1 (PDCD1) and lymphocyte activation gene 3 (LAG3) in γδ T cells. The exogenous IL-35 stimulated group also showed impairment of cytotoxic cytokine secretion. In addition, stat5a revealed a significant increase after IL-35 stimulation of γδ T cells screened via transcription factor based on PCR array analysis. Furthermore, bioinformatics analysis revealed that stat5a-related tumor-specific genes were mainly involved in immune regulatory pathways. Correlation analysis indicated that stat5a expression was significantly and positively correlated with tumor immune cell infiltration, and pdcd1 and lag3 expression. Finally, bioinformatics analysis using the TCGA and GSE36376 HCC datasets corroborated the significant positive correlation between IL-35 and stat5a. Taken together, overexpressed IL-35 promoted exhaustion and impaired the anti-tumor function of γδ T cells in HCC. Targeting IL-35 might be a promising means to enhance the antitumor therapy of γδ T cells, which would significantly improve prognosis.

Violation of the vaginal microbiome and the risk of HIV infection in women

Women are the main population responsible for new HIV infections and the persistence of the HIV pandemic. The article provides a review of the current literature on the factors and mechanisms by which the vaginal microbiome can contribute to HIV infection. A key determinant of susceptibility to HIV infection is the composition of the vaginal microbiome, which can influence the local immune cell population and inflammation status. A low diversity microbial composition dominated by Lactobacillus crispatus does not increase the risk of HIV infection, and a high microbial diversity environment associated with bacterial vaginosis increases the risk of HIV infection. The female reproductive tract has a unique susceptibility to viral infection because tissue-specific immunity must elicit rapid antimicrobial responses to pathogens while maintaining sperm tolerance. It is important to note that the risk of HIV infection during sexual intercourse is multifactorial and is determined not only by the state of the microbiota of a woman’s genital tract, but also by the state of the partner’s microbiota, the viral load of HIV in the blood of the sexual partner, the presence/absence of anti-retroviral therapy in the partner, hormonal background and the phase of the menstrual cycle, which also affect the state of the microbiota.

Changes in cervical cytokine profiles following focused ultrasound treatment for high-risk human papillomavirus infection-related low-grade cervical lesions.

To evaluate the changes of cervical soluble immune markers after focused ultrasound (FU) treatment to explore the underlying local immune effects of FU in the treatment of high-risk human papillomavirus (HR-HPV) infection-related low-grade squamous intraepithelial lesion (LSIL). A total of 35 patients diagnosed with HR-HPV infection-related histological LSIL who met the inclusion criteria were enrolled in this prospective study and treated with FU. The authors used cytometric bead array to measure T-helper type 1 (Th1) cytokine (interleukin [IL] 2, tumor necrosis factor, and interferon γ) and Th2 cytokine (IL-4, IL-5, IL-6, and IL-10) levels in the cervicovaginal lavage of patients before and 3 months after FU treatment. After FU treatment, the concentrations of Th2 cytokines IL-5 and IL-6 were significantly lower than those before FU treatment (P = 0.044 and P = 0.028, respectively). HR-HPV infection was cleared in 27 patients, with a clearance rate of 77.1% (27 of 35). The concentration of IL-4 in patients with HR-HPV clearance after FU treatment was significantly lower than that in patients without HR-HPV clearance (P = 0.045). FU can inhibit the production of certain Th2 cytokines and may improve the local immune status of the cervix, thereby eliminating HR-HPV infection.

Morphopathogenesis of Adult Acquired Cholesteatoma.

Background and Objectives. The aim of this study was to compare the distribution of proliferation markers (Ki-67, NF-κβ), tissue-remodeling factors (MMP-2, MMP-9, TIMP-2, TIMP-4), vascular endothelial growth factor (VEGF), interleukins (IL-1 and IL-10), human beta defensins (HβD-2 and HβD-4) and Sonic hedgehog gene protein in cholesteatoma and control skin. Methods. Nineteen patient cholesteatoma tissues and seven control skin materials from cadavers were included in the study and stained immunohistochemically. Results. Statistically discernible differences were found between the following: the Ki-67 in the matrix and the Ki-67 in the skin epithelium (p = 0.000); the Ki-67 in the perimatrix and the Ki-67 in the connective tissue (p = 0.010); the NF-κβ in the cholesteatoma matrix and the NF-κβ in the epithelium (p = 0.001); the MMP-9 in the matrix and the MMP-9 in the epithelium (p = 0.008); the HβD-2 in the perimatrix and the HβD-2 in the connective tissue (p = 0.004); and the Shh in the cholesteatoma's perimatrix and the Shh in the skin's connective tissue (p = 0.000). Conclusion. The elevation of Ki-67 and NF-κβ suggests the induction of cellular proliferation in the cholesteatoma. Intercorrelations between VEGF, NF-κβ and TIMP-2 induce neo-angiogenesis in adult cholesteatoma. The similarity in the expression of IL-1 and IL-10 suggests the dysregulation of the local immune status in cholesteatoma. The overexpression of the Sonic hedgehog gene protein in the cholesteatoma proves the selective local stimulation of perimatrix development.