[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 significance of interleukin-6 and polymorphism of its gene (C174G) under viral, alcoholic and mixed cirrhosis of liver].

The study was carried out to evaluate significance of interleukin-6 (IL-6) and polymorphism of gene IL-6 (C174G) under liver cirrhosis of viral, alcoholic and mixed etiology. The blood of patients with liver cirrhosis of viral (n=48), alcoholic (n=11) and mixed (n=10) genesis was used for analyzing indices, level of IL-6 and polymorphism of gene IL-6 (C174G). In patients with liver cirrhosis independently of etiology increasing of concentration of IL-6 was established as compared with control group (p<0.001). The gene reliably correlated with marker of cytolysis (r=0.32; p=0.009), direct bilirubin (r=0.26; p=0.04), gamma-glutamyltranspeptidase (r=0.36; p=0.004), albumin (r=0.59; p<0.001) and degree of severity of liver cirrhosis according the Child-Pugh scale (r=0.76; r<0.001). The concentration of IL-6 was significantly higher in patients with alcoholic liver cirrhosis tahn in patients with viral and mixed genesis of disease (p=0.02 and p=0.02 correspondingly). The analysis of prevalence of genotypes and alleles of polymorphism of gene IL-6 (C174G) in groups of donors and patients with liver cirrhosis in the Permskii Kraii established no reliable differences in rate of occurrence of minor allele G and its association with production of IL-6 in examined cohorts (p=0.82 and p=0.29 correspondingly). This result indicates absence of relationship of probability of immune disorders and their genetic determination. The development of liver cirrhosis independently of etiologic factor is characterized by hyper-production of anti-inflammatory cytokine reflecting severity of lesion of hepatocytes, intensity of inflammatory process and progression of disease. The more intensive production of IL-6 was observed in case of alcoholic genesis of disease that conditioned fast progression of cirrhosis.

[The laboratory evaluation of dynamics of progression of liver fibrosis under chronic hepatitis].

The study was carried out to evaluate velocity of development of fibrosis of liver in patients with chronic hepatitis C using laboratory tests. The sampling of 150 patients with chronic hepatitis C were examined to analyze blood serum in phase of reactivation detecting concentration of hyaluronic acid, alpha-fetoprotein, iron, ferritin, vascular endothelial risk factor, malonic dialdehyde and activity of catalase. The degree of fibrosis of liver was evaluated using technique of ultrasound elastography. The velocity of development of fibrosis of liver was calculated as ratio of fibrosis (in points) to duration of infection (in years). The velocity of development of fibrosis of liver in group with slow rate of progression of disease made up in average 0.01±0.04 points per year and in the group with fast rate of progression of chronic hepatitis C - 0.45±0.34 points per year (p<0.001). In the group of patients with high velocity of development of fibrosis of liver were registered reliably higher values of density of liver (p<0.001). The fast velocity of development of fibrosis of liver was accompanied by increasing of serum concentrations of hyaluronic acid (p<0.001), alpha-fetoprotein (p=0.04), iron (p=0.03), ferritin (p=0.03) and decreasing of activity of catalase (p=0.04). The correlation analysis substantiated availability of reliable direct relationships of rate of progression of fibrosis with duration of disease (r=0.27, p=0.01), density of liver (r=0.86; p<0.001), hyaluronic acid (r=0.59; p<0.001), alpha-fetoprotein (r=0.23; p=0.04), ferritin (r=0.23; p=0.04), vascular endothelial risk factor (r=0.21; p=0.04). The association of increased serum levels of hyaluronic acid, alpha-fetoprotein, vascular endothelial risk factor, iron, ferritin and low activity of catalase with fast velocity of development of fibrosis of liver can be recommended as additional criteria of evaluation of rate of progression of fibrosis under chronic hepatitis C.

[The dysfunction of endothelium and humoral mediators of blood plasma in patients with ulcerous colitis.]

The study was carried out to evaluate biological reaction of endothelium-depended vasodilation and disorder of its regulation by humoral mediators of inflammation in patients with ulcerous colitis. And also to compare disorders of function of endothelium with content of cytokines in blood plasma and severity of inflammation in intestine. The examined sampling consisted of 90 patients with diagnosis of ulcerous colitis at the phase of exacerbation of disease. The concentration of humoral mediators of inflammation was established: tumor necrosis factor-alpha, interleukins-6 and -4, monocyte chemo-attractant protein-1. The content of endothelium growth factor of vessels in blood serum was determined as a marker of damage of endothelium. The functional condition of endothelium of vessels was evaluated on the basis of photoplethysmography and software support Pulsware (Samara). In case of ulcerous colitis hyper-secretion of humoral mediators of inflammation was established in blood serum: tumor necrosis factor-alpha, interleukins-6, monocyte chemo-attractant protein-1. The median of concentration of endothelium growth factor of vessels thrice exceeded the level specific for control group of healthy individuals. The disorder of biological reaction of endothelium-depended vasodilation was established in 95% of patients with ulcerous colitis. The reliable correlation relationship between concentration of humoral mediator of inflammation, endothelium growth factor of vessels and severity of destructive alterations in large intestine wall. The combination of disorders of humoral regulation of biological reaction of endothelium-depended vasodilation and markers of dysfunction of endothelium reflects activity of local destruction of tissue. The severity of clinical manifestation of ulcerous colitis substantiates functional interaction of inflammation reaction and angiogenesis. The detection of humoral mediators of inflammation and endothelium growth factor of vessels is reasonably to implement as non-invasive techniques of evaluation of severity of inflammation in wall of large intestine under ulcerous colitis.

[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.

[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.

[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.

[The noninvasive evaluation of degree of expression of fibrosis of liver and significance of polymorphism of gene of hyaluronic acid under chronic hepatitis C].

The study was carries out to evaluate degree of expression of fibrosis, reparation processes in liver and value of polymorphism of gene of hyaluronic acid HASI (rs11084111) in progression of affection of liver in patients with chronic hepatitis C. The sampling included 100 patients with chronic hepatitis C. The control group included 83 healthy donors. The blood serum was tested to detect concentration of hyaluronic acid and alpha-fetoprotein. The stage of liver fibrosis (F) was evaluated by using ultrasound fibroflexography The polymorphism of gene (rs11084111) was analysed by polymerase chain reaction technique. In the group of patients with F1 the average concentration of hyaluronic acid in blood serum in 1.8 times surpassed this indicator in group with F0. The concentration of hyaluronic acid was almost 2 times higher under F3 as compared with F1-F2. This indicator permitted differentiating F3 and F4 which followed by activation of cytolysis and cholestasis in F1 and F3 and by increasing of level of alpha-fetoprotein at stages F1 and F4. The study detected no statistically significant difference between rates of genotypes and alleles of gene HASI (rs11084111) in groups of healthy patients and patients with chronic hepatitis C. The direct relationships are established between hyaluronic acid and markers of cytolysis, cholestasis, alpha-fetoprotein (p = 0.001), viral load (p = 0.003) liver elasticity index according fibroflexography data (p < 0.001) and fibrosis index (p < 0.001). The established relationships indicate association of hepatofibrosis with cytolysis, cholestasis, hepatocytes regeneration and virus activity. The hyaluronic acid permits to stratify minimal expressed fibrosis and also the transition of disease to the stage of cirrhosis.

[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.

[The intramural neural apparatus of the mesenteric lymphangion of the small intestine]. / Intramural'nyi nervnyi apparat limfangiona bryzheiki tonkoi kishki.

Concentration of nerve structures in the lymphangion wall directly correlates with myocyte number in its different regions. Maximal adrenergic and cholinergic fibres concentration was defined in valvular torus muscular cuff of the lymphangion and minimal--in the valvular sinus wall. Besides, as structural organization of adrenergic plexus does not differ from that of antichollinaestherase-positive one and electron microscopic photos display diverse cholinergic and monoaminergic mediator vesicles in same fibres a suggestion is made on the mixed mediator nature of intramural plexuses of the lymphangion.