[Method of diagnosis of the severity of the disease attack in patients with ulcerative colitis.]

To develop a mathematical model and evaluate its prognostic significance for determining the severity of attack (SA) in patients with ulcerative colitis (UC) on the basis of minimally invasive laboratory inflammatory tests and albumin. The study involved 64 patients (33 men and 31 women) with UC in the stage of active inflammation. The method for diagnosing SA was performed by determining the concentration in the blood of tumor necrosis factor alpha (TNF-α), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), α2-globulin and albumin. Truelove and Witts criteria (1955) were used as a reference method for the evaluation of SA in patients with UC. In constructing a mathematical model, the severity of the attack of the UC emerged as a dependent variable. As independent variables or predictors - five laboratory parameters: TNF-α, CRP, ESR, α2-globulins and albumin, which had a high relationship with SA. At SA value in the range from 0 to 0.5, absence of attack or remission of the disease is determined, SA value in the range from 0.5 to 1.2 corresponds to mild severity of UC attack, SA value in the range from 1.2 to 2.2 corresponds to the moderate severity of the attack of the UC, at SA more than 2.2 one should speak of a severe attack of the UC. The model is statistically significant: the Fisher test F = 439.9; p < 0.0001, multiple R = 0.981; R2 = 0.961. The diagnostic sensitivity of this mathematical model was 98.4%, specificity - 96.7%. The index of the severity of the attack, calculated by the formula proposed by us, is able to differentiate the mild, moderate and severe SA, including the isolation of patients with a stage of remission. In this case, the method is non-traumatic, has a low cost and high sensitivity and specificity.

[The non-invasive accessible informative laboratory panel for detection of liver fibrosis - TEA index.]

The article presents the results of continued studies of informative accessible tests as a screening non-invasive technique of diagnostic of fibrosis under chronic hepatitis C. The sampling included 70 patients with chronic hepatitis C. The comparable control group included 30 healthy individuals. The ultrasonic elastography of liver was implemented using FibroScan («EchoSens¼, France). The concentration of tumor necrosis factor-alpha (TNF-α) in blood serum was detected by the enzyme-linked immunosorbent assay technique («StatFax¼, USA) using reagents of "Vektor-Best" (Russia); thrombocytes - by hematologic analyzer Medonic-620M (Sweden); albumin - by analyzer Archtekt-4000. The ROC-analysis and detection of odds ratio (QR) was implemented to calculate threshold values and diagnostic efficiency of indices with predictor value. The correlation relationship is established between density of hepatic tissue according ultrasonic elastography data and three applied tests: number of thrombocytes (r = -0,9), content of TNF-α (r = 0,89) and albumin (r = -0,9). These are the three tests included into mathematical model of diagnostic of fibrosis stage. The equation of multiple regression is reproduced in the utility patent № 2592371. The diagnostic value of the proposed index of fibrosis TTA (thrombocytes, TNF and albumin) according the scale METAVIR for F0 made up to 0-0.5; for F1-2 - 0.6-2.5; in case of expressed fibrosis/cirrhosis index made up to more than 2.5. The index TTA by its efficiency is comparable with more complicated analogues. The individual application of fibrosis predictors is possible: the test permit to exclude fibrosis under chronic hepatitis C at number of thrombocytes in blood more or equal to 282×109l,, value of TNF-α les or equal 1.9 pg/ml, level of albumin more or equal 47.3 g/l and also to differentiate stages of moderate (F1-2) and expressed fibrosis (F3-4). The index TTA of liver fibrosis can be applied to exclude fibrosis under chronic hepatitis C and also to establish stages of fibrosis with diagnostic sensitivity 93.3% and specificity 83%. At that, low approximate cost of examination has an important value.

[THE INFLUENCE OF TUMOR NECROSIS FACTOR ALPHA AND POLYMORPHISM OF ITS GENE (RS1800629) ON THE SEVERITY AND PROGRESSION OF CHRONIC HEPATITIS AND ULCERATIVE COLITIS].

AIM OF THE STUDY: To investigate the effect of generation of tumor necrosis factor-alpha (TNFα) and the importance of TNFα(rs1 800629) gene polymorphism in the progression of chronic hepatitis C (CHC) and ulcerative colitis (UC). MATERIAL AND METHODS: The study involved 90 patients with chronic hepatitis C, 50 patients with UC and 50 healthy donors. The blood concentrations TNFα and TNFα gene polymorphism (rs1800629) were evaluated. RESULTS: TNFα levels in the blood in patients with chronic hepatitis C were increased compared with the control group and correlated with the severity of Cytolysis and fibrosis (r = 0.34, p = 0.02). At slow speed the formation of liver fibrosis TNFα amounted to 1.5 (0.9-2.8) pg/mI, with a fast speed--2.3(1.4-8.2) pg/mI (p = 0.006). Patients with UC at 3-4 degrees endo- scopic activity production of TNFα reached 6.5 (7-9) pg/mI, which was significantly higher than the value obtained at 1-2 degrees endoscopic activity--0.25(0-0.8) pg/ml (p = 0.001). The allelic variations of TNFα in groups of patients with CHC at different rates forming LF statistically differences were not found. The allele, associated with severe progressive course of UC and increased production of TNFα--A risk allele and genotype GA TNFα, associated with a slow progression of UC- "protective" G allele and genotype GG TNFα gene were determined. CONCLUSION: Determining the level of TNFα allows to evaluate the severity of liver disease, heaviness and progression of liver fibrosis speed in CHC, and the severity of inflammation in the intestinal mucosa in UC. The presence of the allele A of TNFo(rs1800629) is a predictor of severe and progression of UC. Determining genetic polymorphism TNFα in patients with UC may be an additional factor to assess the prognosis of the disease.

[Metabolic disorder and polymorphism of the genes encoding for beta-2-adrenergic receptor and apolipoproteins B in chronic hepatitis C and non-alcoholic fatty liver diseases].

UNLABELLED: The aim of the study was to evaluate the relationship between metabolic disorders and polymorphic variants of the genes encoding for beta-2-adrenergic receptor (ADRB2 (rs1042713) and apolipoproteins B (ApoB(rs5742904) in patients with chronic hepatitis C (CHC) depending on virus genotype and in patients with non-alcoholic fatty liver diseases (NAFLD) and concomitant metabolic syndrome (MS). MATERIALS AND METHODS: The study included 96 patients with CHC (51 with genotype 1 or 2 of hepatitis virus and 45 with genotype 3), 70 patients with NAFLD and MS, 51 healthy donors (controls). Gene polymorphism was studied by PCR (Sintol, Moscow) with a Real-time CFX-96 amplifier (Bio-Rad Lab. Inc., USA). RESULTS: CHC patients regardless of genotype had hypertriglyceridemia with increased atherogenicity index and C-peptide level. Hyperleptinemia was most frequently associated with genotype 3, hyperinsulinemia and insulin resistance with genotypes 1 and 2. The viremia level positively correlated with leptin level (p-0.021) and HOMA-IR index (p=0.022) which suggests virus-induced inactivation of mechanisms of steatogenesis and insulin resistance. Leptin production in CHC was associate with activation of liver fibrosis as appears from elasticity index measured by fibroelastography (p-0.22). Almost all patients with NAFLD had disturbances of lipid and carbohydrate metabolism. Hepatic lesions in 30% of the patients with MS were accompanied by laboratory signs of steatohepatitis. Patients with CHC showed an increased frequency of minor A allele of the ADRB2 (rs1042713) gene (up to 40%; p=0.04) and pathological A/A homozygote (22%; p=0.04). The occurrence of A allele was associated with hyperleptinemia (p=0.019). In NAFLD patients, the occurrence of A allele was higher than in controls (41%; p=0.02) with 55% of the case being A/G heterozygotes (p=0.005). The occurrence of A allele was related to hyperinsilinism (p=0.036), BMI (p=0.0330, and C-peptide production (p=0.038). No difference between the frequency of genotypes and ApoB(rs5742904) gene alleles in the patients of both groups and controls was documented. It is concluded that ADRB2 (rs1042713) gene polymorphism is associated with an increased risk of metabolic disorders in CHC and especially NAFLD with MS that aggravates liver disturbances. Dyslipidemia and insulin resistance in CHC patients is stimulated by viral infection.

[Catalase gene rs1001179 polymorphism and oxidative stress in patients with chronic hepatitis C and ulcerative colitis].

AIM: To study the rs1001179 polymorphism of the catalase (CAT) gene and to estimate the serum levels of the enzymes catalase and glutathione peroxidase (GP) in patients with chronic hepatitis C (CHC) and in those with ulcerative colitis (UC) in the Perm Territory. SUBJECTS AND METHODS: Ninety patients with reactivation-phase CHC and 50 patients with exacerbation-phase UC were examined. The serum levels of catalase and GP were determined and the polymorphic variants of the marker of CAT gene rs1001179 in the DNA isolated from whole blood were found in all the patients. RESULTS: In the CHC and UC groups, the levels of catalase and GP were found to be lower than that in apparently healthy individuals. Furthermore, both groups showed a direct correlation between the activities of the enzymes. In the patients with CHC and in those with UC, the spread of genotypes and alleles generally failed to virtually differ from that in the control group. The G/G genotype was prevalent in all the groups. In the patients with CHC, the minor A allele demonstrated a significant inverse correlation with the enzyme catalase (r = -0.16; p = 0.02) and GP (r = -0.13; p = 0.047). CONCLUSION: The lower serum levels of catalase and GP are indicative of oxidative stress in the patients with CHC or UC. In the patients with CHC, the significant correlation of the pathological rs1701179 A allele marker with the processes of synthesis of antioxidant enzymes may suggest that CAT gene polymorphism in the A/A homozygotes might affect the regulation mechanism involved in the antioxidant system in the liver.

[The relationship between content of malonic aldehyde, activity of glutathione peroxidase and polymorphism of gene GPX4 (718C/T) under chronic hepatitis C].

The study was carried out to evaluate concentration of malonic dialdehyde and activity of glutathione peroxidase in blood serum in their relationship with biochemical tests of functional conditions of liver; data of fibroelastography and polymorphism of gene GPX4 (718C/T) in patients with chronic hepatitis C. The sampling consisted of 100 patients with chronic hepatitis C. The control group included 80 healthy donors. The polymorphism of gene of glutathione peroxidase was analyzed with CFX-96 (Bio-Rad Laboratories, Inc., USA) amplifier using technique of polymerase chain reaction ("Sintol", Moscow). In patients with chronic hepatitis C a reliable increasing of concentration of malonic dialdehyde up to 3.2 times and decreasing of glutathione peroxidase up to 2.8 times was detected as compared with control group. The significant differences in rates of genotypes and alleles of gene GPX4 (718C/T) in patients with chronic hepatitis C and healthy persons were not detected. The malonic dialdehyde demonstrated direct reliable relationships with functional hepatic tests by activity of alanine aminotransferase, aspartate aminotransferase and γ-glutamyl transferase. The activity of glutathione peroxidase had reverse reliable correlations with alanine aminotransferase, aspartate aminotransferase and γ-glutamyl transferase (p=0.001, p=0.007, p=0.032) and with indicator of elasticity of liver according data of fibroelastography (r=-0.285, p=0.041) The minor allele T of gene GPX4 ()718C/T) reliably correlated with alanine aminotransferase, aspartate aminotransferase and malonic dialdehyde. The reverse relationship between allele T of gene GPX4 ()718C/T) and activity of glutathione peroxidase (r=-0.196, p=0.041) was established This occurrence indicates at negative effect of mutation in gene of glutathione peroxidase on functional activity of this enzyme. Under chronic hepatitis C, the activation of peroxidation of lipids and depression of activity of glutathione peroxidase are interrelated with severity of cytolysis, cholestasis, expression of hepatic fibrosis and polymorphism of gene GPX4 ()718C/T). Therefore, polymorphism of gene of glutathione peroxidase predisposes to more severe affection of liver and progression of chronic hepatitis C.

[Diagnostic efficiency of laboratory tests determining the functional status of the endothelium in patients with chronic diffuse liver diseases].

The patients with chronic diffuse liver diseases were found to have signs of endothelial damage, which manifests itself as increases in the count of desquamated endotheliocytes, the level of vascular endothelial growth factor, and the concentration of von Willebrand factor in plasma and to have signs of endothelial dysfunction as reduced nitric oxide levels and elevated endothelin-1 concentrations in plasma. The magnitude of changes in the indicators of endothelial dysfunction and in the markers of endothelial damage depends on the severity of hepatic damage.