Blood Level of Nitric Oxide Metabolites and Expression of Genes Regulating NO Synthesis in Early Forms of Non-Alcoholic Fatty Liver Disease.

The levels of NO metabolites in the plasma and mRNA of the NOS3, ATG9B, and NOS2 genes in peripheral blood leukocytes of healthy people and patients with early forms of non-alcoholic fatty liver disease (steatosis and weak activity non-alcoholic steatohepatitis) were studied. In patients with steatohepatitis, the concentration of NO metabolites in the blood and the level of mRNA of the NOS2 gene were higher than in patients with steatosis and healthy people. These differences can be of diagnostic value for distinguishing between steatosis and weak activity steatohepatitis in non-alcoholic fatty liver disease. A correlation between the levels of NO metabolites and the expression of the NOS2 gene in weak activity steatohepatitis was established, which indicates activation of NO synthesis in non-alcoholic steatohepatitis due to the expression of the inducible NO synthase gene. The level of the NOS2 gene mRNA in peripheral blood leukocytes of patients with weak activity steatohepatitis correlated with the level of TNFα and IL-6 cytokines. An increase in the level of NO in the blood in weak activity steatohepatitis correlated with the level of MDA, an indicator of oxidative stress.

The Role of the Soluble Interleukin-6 Receptor in the Progression of Nonalcoholic Fatty Liver Disease.

The blood level of soluble IL-6 receptor was measured in patients with different clinical and morphological forms of nonalcoholic fatty liver disease and healthy donors. The relationship of the soluble IL-6 receptor with the content of IL-6, the level of the IL6 gene mRNA, and a number of markers of hepatocyte and peripheral blood leukocyte apoptosis was assessed. It has been established for the first time that progression of nonalcoholic fatty liver disease is associated with changes in the level of soluble IL-6 receptor in the blood. In patients with high activity of nonalcoholic steatohepatitis and liver cirrhosis, the blood concentration of soluble IL-6 receptor sharply decreased in comparison with the earlier stages of progression of nonalcoholic fatty liver disease (liver steatosis, nonalcoholic steatohepatitis of weak and moderate activity). This allows considering the decrease in this indicator as a new diagnostic marker for distinguishing nonalcoholic steatohepatitis of high activity from weak and moderate activity. A close correlation between changes in the level of soluble IL-6 receptor and apoptosis of peripheral blood leukocytes and hepatocytes was revealed.

Hepatocytic Apoptosis and Immune Dysfunction in Decompensation of Alcoholic Liver Cirrhosis with Different Grades of Acute-on-Chronic Liver Failure.

Acute-on-chronic liver failure (ACLF) of varying grades was assessed in 110 patients with alcoholic liver cirrhosis using the on-line CLIF-C ACLF Calculator ( www.efclif.com/scientific-activity/score-calculators/clif-c-aclf ); fragments of cytokeratin-18, TNFα, IL-1ß, IL-4, IL-6, and IL-8 were also assayed. As ACLF progressed from grade 0 to grade 3, the levels of cytokeratin-18 fragments, IL-6, and IL-8 significantly increased, while IL-4 decreased. TNFα peaked in ACLF grade 1, but decreased in grades 2 and 3. IL-1ß did not depend on the ACLF grade. Thus, hepatic damage and immune dysfunction are implicated in the progression of ACLF.

Role of Bacterial Infection in the Development of Acute Liver Failure in Patients with Decompensated Alcoholic Liver Cirrhosis.

We examined 74 patients with acute decompensation of alcoholic liver cirrhosis: 34 (45.9%) with bacterial infection (group 1) and 40 (54.1%) without bacterial infection (group 2). The degree and index of acute-on-chronic liver failure (ACLF) were determined using an on-line CLIF-C ACLF Calculator and the levels of cytokeratin-18 fragments, TNFα, IL-1ß, IL-4, IL-6, and IL-8. In group 1, AST, cytokeratin-18, TNFα, IL-1ß, IL-6, degree and score of ACLF were significantly higher than in group 2. ACLF developed in 18 (52.9%) patients in group 1 and in 11 (27.5%) (p<0.05) patients in group 2. Within 1 month, 10 (29.4%) patients of group 1 and 2 (5%) patients of group 2 died (p<0.05). Patients with bacterial infection showed a more severe course of alcoholic liver cirrhosis and ACLF than those without bacterial infection.

[Hepatocellular damage and inflammation in various forms of alcoholic liver disease].

AIM: The aim of the study was to evaluate hepatocellular damage and immune inflammation in various forms of alcoholic liver disease (ALD). MATERIALS AND METHODS: 104 patients with ALD were examined: 15 (14.4%) with liver steatosis (LS), 19 (18.3%) with steatohepatitis and 70 (67.3%) with liver cirrhosis (LC); men 50 (48.1%), women 54 (51.9%); age 45.78.4 years. Traditional clinical, laboratory, instrumental studies were performed, the levels of fragments of cytokeratin-18 (FCK-18), cytokines IL-1, TNF-, IL-4, IL-6, IL-8 were determined by ELISA. The control group consisted of 39 healthy individuals: men 20 (51.2%), women 19 (48.7%), age 48.58.3 years. RESULTS: In LS, an increase in the level of FCK-18 was noted with normal aminotransferase activity, the content of TNF-, IL-6, IL-1, IL-8 increased and the level of IL-4 decreased compared to those in healthy individuals. In steatohepatitis, a triple increase in aminotransferases and FCK-18 was observed compared with LS, as well as an increase in the level of inflammatory mediators, to a greater extent IL-6, to a lesser extent IL-8, TNF-, a decrease in IL-4, IL-1 remained at the same level. In LC, there was a further increase in FCK-18, significantly more pronounced than an increase in AST, and the increase in cytokines continued to the same extent, the levels of IL-6 and IL-8, to a lesser extent IL-1 and TNF-, and the level of IL-4. CONCLUSION: With the progression of ALD from LS to steatohepatitis, hepatic cell damage was carried out by equally pronounced processes of hepatocyte necrosis and apoptosis, with the development of cirrhosis of the liver, parenchyma damage occurred mainly due to hepatocyte apoptosis. The immuno-inflammatory process progressively increased from the stage of LS to LC with IL-6 and IL-8 undergoing the greatest dynamics. FCK-18 can serve as a non-invasive marker of hepatic cell damage, and IL-6 and IL-8 markers of immune inflammation in ALD.

Biochemical and molecular-genetic indicators of inflammation and apoptosis in liver cirrhosis as an outcome of the progression of non-alcoholic steatohepatitis.

AIM: A comparative analysis of the complex of clinical and laboratory indicators (including the content of cytokines in blood plasma and the level of expression of TNF and IL6 genes in peripheral leukocytes, as well as the level of biochemical and molecular-genetic indicators of apoptosis, such as the content of tissue polypeptide-specific antigen (TPS) in the blood, the activity of caspases 3, 8 and 9 and the expression level of the encoding genes in peripheral blood leukocytes) in patients with non-alcoholic fatty liver disease (NAFLD) with non-alcoholic steatohepatitis (NASH) of different activity, liver cirrhosis (LC) classes A and B and in the donors of control group. MATERIALS AND METHODS: 158 patients with NAFLD were examined: 116 patients with NASH diagnosed for the first time (NASH of weak, moderate and high activity) and 42 patients with the NAFLD at the stage of liver cirrhosis diagnosed for the first time (classes A and B according to the Child-Pugh classification). The control group consisted of 54 healthy donors. The clinical blood biochemistry, cytokine profile, tissue polypeptide-specific antigen content, the level of the TNF, IL6 gene and caspase gene transcription as well as caspase activity in peripheral blood leukocytes (PBL) were evaluated. RESULTS: In the progression of NASH to LC, together with changes in general clinical parameters, the cytokine profile are changed due to an increase in the level of IL-6 and IL-1ß; in peripheral leukocytes, the activity of caspase 9 increases and the activity of caspase 8 decreases compared to NASH, and the level of the TNF gene expression decreases as compared to NASH of high activity. These parameters can be considered as promising minimally invasive markers of progression of NAFLD to LC. CONCLUSION: In nonalcoholic cirrhosis as an outcome of the progression of non-alcoholic steatohepatitis changes in clinical parameters (indicating the development of hepatocellular deficiency, violation of protein and lipid metabolism, progressive inflammation) are accompanied by specific changes in levels of biochemical and molecular-genetic indicators of apoptosis and inflammation. With the progression of NASH to LC, the cytokine profile changes due to an increase in the level of proinflammatory cytokines, the apoptosis processes triggered by the internal pathway increase and the activity of apoptosis activated via the external pathway decreases in PBL.

IL6R Gene Polymorphic Variant rs2228145(C >A) as a Marker of Genetic Liability to Nonalcoholic Steatohepatitis in the Russian Population of Karelia.

Association of IL6R gene polymorphic variant rs2228145(C>A) with the development of nonalcoholic steatohepatitis in Karelia residents is detected. The risk of nonalcoholic steatohepatitis is more than 2-fold higher in carriers of CC genotype by rs2228145 polymorphic marker than in carriers of other genotypes. Plasma levels of IL-6 and the content of IL6R gene transcripts in the peripheral blood leukocytes are higher in patients with nonalcoholic steatohepatitis than in normal subjects. No relationships between rs2228145 polymorphism and the level of IL-6 and content of IL6 and IL6R mRNA were detected. Gene IL6R polymorphic variant rs2228145(C>A) seems to be involved in genetic predisposition of the population of Karelia to nonalcoholic steatohepatitis. However, biochemical and molecular mechanisms underlying the relationship of rs2228145 with the development of nonalcoholic steatohepatitis are not yet studied.

The clinical significance of insulin resistance in non-diabetic patients with early forms of non-alcoholic fatty liver disease.

AIM: To assess the presence of insulin resistance (IR) in non-diabetic patients with early forms of non-alcoholic fatty liver disease (NAFLD) - liver steatosis (LS) and steatohepatitis (SH) of mild activity and the influence of IR on the clinical course of these diseases. MATERIALS AND METHODS: 134 patients with NAFLD were examined: 54 with LS and 80 with SH. The control group consisted of 37 healthy donors. Anthropometric parameters (body mass index (BMI), waist circumference (WC)), clinical and biochemical blood indices, including the blood level of cytokeratin-18 fragments (CK-18), TNF-α and IL-6 cytokines, insulin were evaluated. The HOMA index and the fibrosis index (NAFLD FS) were calculated. Patients were divided into groups: I - with the absence of IR (HOMA-index <2.7), II - with the presence of IR (HOMA-index> 2.7). RESULTS: Indicators of hepatic injury, inflammation, cholestasis, fibrosis and atherogenic dyslipidemia are higher in patients with LS of group II (with IR) than in group I patients (without IR). BMI, WC, γ-glutamil transpeptidase, CK-18 and fibrosis index are significantly higher in group II patients with SH compared with group I, there is no significant difference in the level of cytolysis, inflammation and dyslipidemia indices. A high incidence of IR in non-diabetic patients with LS (37.0%) and SH (55.0%) was found and the effect of IR on the clinical course of these diseases was revealed. CONCLUSION: Insulin resistance in non-diabetic patients with NAFLD was detected in SH (55.0%) with higher frequency than in LS (37.0%). In LS, IR is associated with impaired hepatic cell damage, intrahepatic cholestasis, atherogenic dyslipidemia and fibrosis. In SH, IR is combined with reliable growth in indicators of hepatocyte apoptosis, cytokine proinflammatory status and fibrosis. IR determines the progressing course of NAFLD, promoting the transformation of steatosis into steatohepatitis and steatohepatitis into fibrosis and liver cirrhosis.

Intrahepatic cholestasis in nonalcoholic fatty liver disease.

AIM: To determine the frequency of intrahepatic cholestasis and its impact on the clinical features of the different forms of non-alcoholic fatty liver disease (NAFLD). MATERIALS AND METHODS: The study involved 163 patients with NAFLD: 92 (56.4%) with hepatic steatosis (HS), 56 (34.4%) with steatohep- atitis (SH) and 15 (9.2%) with liver cirrhosis (LC). Diagnosis is based on clinical, laboratory, ultrasound and histological data. Insulin, tu- mor necrosis factor α (TNF-α), fragments of cytokeratin-18 (FCK-18) were determined by ELISA. The index of insulin resistance (HOMA- IR) was calculated. NAFLD fibrosis score (NAFLD-FS) was determined, taking into account the patient's age, body mass index, presence or absence of carbohydrate metabolism disturbances, levels of ASAT, ALAT, albumin and blood platelets. RESULTS: Cholestatic syndrome was detected in 49 (30.1%) NAFLD patients: in 23 (25%) with HS, in 19 (33.9%) with SH and in 7 (46.7%) with LC. Patients with HS, SH and LC with signs of cholestasis as compared to patients with the same forms of NAFLD without cholestasis had significantly higher levels of the following indicators: aminotransferases, triglycerides, HOMA-IR, TNF-α, FCK-18, NAFLD-FS, - the number of platelets is reduced, indirectly confirming the more rapid development of fibrosis in cholestasis. These findings were consistent with published data on the violation in cholestasis regulatory functions of bile acids, which are ligands of hepatocyte nuclear receptor, re- sponsible for normal homeostasis. CONCLUSION: In all forms of NAFLD with cholestasis were detected more pronounced liver cell inflammation, hepatocyte necrosis and apoptosis, fibrosis, disturbance of carbohydrate and lipid metabolism, which contributed to a progressive course of NAFLD and confirmed the need for medical correction of cholestasis, starting with the earliest form of NAFLD - hepatosteatosis.

Gene TNF Polymorphism -308G>A (rs1800629) and Its Relationship with the Efficiency of Ursodeoxycholic Acid Therapy in Patients with Nonalcoholic Stetohepatitis.

Association of TNF gene polymorphism -308G>A with the development of nonalcoholic steatohepatitis in the Russian population was revealed. Carriers of allele A of the TNF gene marker -308G>A have significantly higher risk of nonalcoholic steatohepatitis development: OR=1.69 (1.05; 2.71). Allele A carriage by this marker predicts an increase in the basal HDL level and a decrease in LDL and IL-10 levels in the blood of healthy subjects. Patients with nonalcoholic steatohepatitis, differing by the TNF gene -308G>A marker genotype, differ by the time course of the markers of hepatocellular damage (ALT, AST), activity of hepatocyte apoptosis (tissue polypeptide-specific antigen), and activation of specific humoral immunity (γ-globulin) in response to therapy with ursodeoxycholic acid in a dose of 10-15 mg/kg over 4-6 weeks. Carriers of allele A of the TNF gene polymorphic marker -308G>A are more sensitive to ursodeoxycholic acid therapy than carriers of GG genotype.

[Features of a necrotic and inflammatory process in different forms of nonalcoholic fatty liver disease]. / Osobennosti nekroticheski-vospalitel'nogo protsessa pri raznykh formakh nealkogol'noi zhirovoi bolezni pecheni.

AIM: To identify the features of development of a necrotic and inflammatory process in different forms of nonalcoholic fatty liver disease (NAFLD), by comparatively analyzing a full set of clinical and laboratory parameters, including the cytokine status and the expression level of enzyme genes controlling the apoptosis of peripheral leukocytes. SUBJECTS AND METHODS: 86 patients with NAFLD, including 8 (9.3%) with hepatic steatosis (HS), 70 (81.4%) with nonalcoholic steatohepatitis (NASH), 40, 19, and 11 with mild, moderate, and high disease activity, respectively, and 8 (9.3%) with liver cirrhosis (LC), were examined. A control group consisted of 34 healthy donors. Clinical and biochemical blood indices, cytokine profile, and the level of caspase gene transcripts in the peripheral blood leukocytes (PBL) were estimated. RESULTS: As compared to the controls, the patients with HS had higher tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6) levels and lower caspase 3, 6, and 8 mRNA in PBL. The concentration of IL-10 in NASH was higher than that in steatosis and positively correlated with the level of proinflammatory cytokines. The levels of TNF-α and IL6 were higher in the patients with NASH than in the controls. Those of C-reactive protein, γ-globulin, IL-6, and cytokeratin-18 fragment increased with the progression of NASH. In the latter, the transcriptional activity of caspase-3 gene decreased relative to the reference value and negatively correlated with the level of proinflammatory cytokines. In the patients with LC, the gene expression profile of caspases in PBL was similar to that in the control group; the level of IL-6 was higher than that in steatosis and NASH, that of IL-1ß was higher than in HS and positively correlated the concentration of IL-6 and the activity of alanine aminotransferase and aspartate aminotransferase. CONCLUSION: The features of a necrotic and inflammatory process were identified in different forms of NAFLD. When the latter progressed, the cytokine profile and gene expression levels of caspases in PBL altered along with a change in the general clinical picture.

Caspase 3, 6, 8, and 9 Gene Expression in Peripheral Blood Leukocytes and Plasma Concentrations of IL-6 and TNF-α in Carriers of Different Polymorphic Marker -174G>C Genotypes of IL6 Gene Associated with the Risk of Nonalcoholic Steatohepatitis.

We revealed an association of IL6 gene -174G>C polymorphism with the development of nonalcoholic steatohepatitis in the Russian population. The risk is significantly higher in carriers of C allele: OR=1.77 (1.04; 3.02). The effects of -174G>C substitution in IL6 gene involving caspase 9 gene transcripts in peripheral blood leukocytes and on blood content of TNF-α in healthy individuals without clinical manifestations of nonalcoholic steatohepatitis were detected. The content of caspase 9 gene transcripts in peripheral blood leukocytes and plasma level of TNF-α were significantly higher in healthy subjects carrying C allele than in carriers of GG genotype. The levels of caspases 3, 6, 8, and 9 gene transcripts in peripheral blood leukocytes and plasma concentrations of TNF-α in patients with nonalcoholic steatohepatitis did not depend on IL6 genotype by -174G

CHRONIC HEPATITIS WITH DOUBLE B/C INFECTION: VIROLOGICAL, CLINICAL, MORPHOLOGICAL CHARACTERISTICS.

AIM: To estimate the r, virological and clinical characteristics of chronic viral hepatitis (CVH) with double B/C infection. MATERIALS AND METHODS: We examined 282 patients with CVH. Genomes of hepatitis B virus (HBV) and hepatitis C virus (HCV) were studied by PCR in blood and liver (AmpliSens HBV and Amplisens HCV Russia), nuclear proteins (HBcorAg HBV and NS3 HCV) were determined by immunohistochemical method (Novocastra, UK), HBVgenome was sequenced by the Sanger method using ABI prism BigDye Terminator v3.1 kits and ABIPRISM 3100 analyzer (AppliedBiosystems, USA). Indices of histological activity (HAI), fibrosis, and portal vein (PV) congestion index (CI) were calculated by formula CI=SBB/LB V where S is P V cross section area in cm2 and LB V - linear blood flow velocity in cm/s (Vivid Pro- 7 apparatus, USA). RESULTS: CVH with double B/C infection was diagnosed in 85 (30.1%) patients including 44.7% with viral genomes and proteins in the live; 42.4% with HCVviremia, and 12.9% with HBJV/HCVviremia. Maximum CVH activity was documented in patients with latent HBV/HCVviremia (ALT 157.2±59.2 U/, HAI 11.6±1.3,fibrosis 2.8±0.7, C1 0.059±0.005); it was minimal inpatients.without viremia (Alt 76.25±63.0 U/I, HAI 6.7+-0.6,fibrosis 1.7±0.5, CI 0.042±0.001;p <0.05). Patients with latent HBV infection had precore/ore and pres/s mutations in HBVgenome and cytoplasmic localization ofHBcorAg. CONCLUSION: Double B/C infection was diagnosed in 30.1% of the patients with CVH dominated by HCV Patients with latent HBVhadprecore/ore and pres/s mutations. The highest intensity of hepatic cellular inflamation,fibrosis, and PV congestion was associated with HBV/HCV viremia and the lowest with intrahepatic localization of both viruses.

[EFFICIENCY OF URSODEOXYCHOLIC ACID APPLICATION IN NONALCOCHOLIC STEATOHEPATITIS].

AIM: to estimate the efficiency of ursodeoxycholic acid (UDHC) in nonalcocholic steatohepatitis (NASH) by analysis of conventional clinical datas, apoptosis and liver perfusion parameters. MATERIALS AND METHODS: UDHC was used as monotherapy in treatment of 92 NASH patients in daily dose 10-15 mg/kg. We have observed 44 (47.8%) males, 48 (52.2%) females, age was 56.8 ± 7.2 years, BMI was 28.4 ± 2.3 kg/m2, waist circumference was 93.8 ± 8.3 cm. Functional liver tests (ALAT, ASAT, alcaline phosphatase--APh, gamma-glutamyltranspeptidase--GGTP), abdominal ultrasonography and dopplerography of liver blood flow, kaspase-3, 6, 8, 9 genes expression in blood leucocytes were estimated. Periods of controls research and UDCA treatment were: 4-8 weeks in 92 patients, 20-24 weeks in 18 (19.6%) patients and 40-48 weeks in 13 (14.1%) patients. RESULTS: Significant positive dynamics of liver functional tests and decrease of kaspase-3, 6, 9 genes expression in blood leucocytes were observed over 4-8 weeks, normalization of liver tests--over 20-24 weeks and significant amelioration of venous and arterial liver perfusion parameters--over 40-48 weeks. CONCLUSION: Ursodeoxycholic acid in daily dose of 10-15 mg/kg in nonalcocholic steatohepatitis caused positive dynamics of cytolytic and cholestasis parameters, leucocytic apoptosis and venous and arterial liver blood flow parameters.

[DIAGNOSIS AND CLINICAL MANIFESTATIONS OF CIRRHOTIC CARDIOMYOPATHY].

UNLABELLED: The aim of the study was to determine the incidence of cirrhotic cardiomyopathy (CCMP), features of its recognition and clinical manifestations. MATERIALS AND METHODS: The study included 102 patients with alcoholic and viral liver cirrhosis (LC) without cardiovascular history, without viremia and signs of acute alcoholic hepatitis. Echocardiography, electrocardiography (ECG) and brain natriuretic hormone (proBNP) level were investigated in all patients. RESULTS: CCMP signs were detected in 65 (63.7%) of the 102 patients examined. All patients showed signs of electrophysiological myocardial abnormalities in ECG, signs of myocardial injury and the presence of heart failure in echocardiography, increase in proBNP. Interval QT length (0.4689 ± 0.012 s.) was significantly (p = 0.0461) higher than that of the control group. Violation of diastolic relaxation of the left ventricle was detected in 36 (55.6%) patients. The average level of proBNP was 540.85 ± 236.43 pg/ml, significantly different from the level of proBNP in healthy individuals--89.45 ± 26.43 pg/ml. The incidence of CCMP increased progressively with the severity of the Child-Pugh class and was 42.4% in patients of A class, 84.6% (p = 0.0132) in patients of B Class and 100% in patients of C class (p = 0.0219). The length of the QT interval, the proBNP level and frequency of diastolic dysfunction increased with increasing of liver cirrhosis severity. CONCLUSION: The frequency of detection of cirrhotic cardiomyopathy was 63.7%. More pronounced CCMP signs were observed in alcoholic liver cirrhosis than in viral. The frequency of detection and severity of cirrhotic cardiomyopathy progres sively increased with the severity of liver cirrhosis. Brain natriuretic peptide was the most sensitive indicator of myocardial damage in liver cirrhosis irrespective of its etiology.

[NONALCOHOLIC FATTY LIVER DISEASE: DIAGNOSTIC, SYMPTOMS, TREATMENT. GUIDELINES WERE APPROVED BY THE XV GASTROENTEROLOGICAL SCIENTIFIC SOCIETY OF RUSSIA IN 2015].

According to the World Health Organization, it is recorded steady growth of the number of chronic liver diseases, including nonalcoholic fatty liver disease (NAFLD), in recent years. As an independent nosological entity, NAFLD is one of the risk factors for cardiovascular diseases, associated with abdominal-visceral obesity, peripheral insulin resistance (IR), and is regarded as the hepatic component of Metabolic syndrome (MS). Nowadays there are no generally accepted national standards for diagnosis and treatment of NAFLD for physicians, gen- eral practitioners, gastroenterologists in Russia. This was the essential reason in their development. The main reason of The guidelines development is the absence of generally accepted national standards for diagnosis and treatment of NAFLD for physicians, general practitioners, gastroenterologists in Russia. These guidelines are based on the global and local data of treatment experience of NAFLD, recently published in reviews, analytical studies in the literature. Guidelines are intended for physicians, general practitioners, gastroenterologists and contain the description of the preferred approaches to the provision of diagnostic, curative and preventive care of patients NAFLD. The quality of recommendations was grading according to the GRADE approach.

[Hepatic parenchimatous damage and perfusion disturbance in the progression of nonalcoholic fatty liver disease].

AIM: To detect early diagnostic criterions of hepatocellular inflammation and portal hypertension for revealing the progressive course of nonalcoholic fatty liver disease (NAFLD). MATERIALS AND METHODS: We have studied 58 patients with NAFLD: 6 with steatosis, 47 with steatohepatitis (SH), 5 with class A liver cirrhosis (LG). Liver biopsy was performed in 24 (41.4%) patients with estimation of histological activity index (HAI) and fibrosis by Brunt method. Abdominal sonography and dopplerography of portal and spleen venous bloodflow with estimation of portal vein congestive index (CI) were performed in all patients. RESULTS: Among conventional laboratory markers of parenchimatous damage gammaglutamiltransferase (GGTF) was increased more frequently--in 77.6% of patients, then alaninaminotransferase (ALAT)--in 62.0% and aspartataminotransferase (ASAT)--in 51.7% of patients. GGTF correlated more closely with HAI: with fatty cellular degeneratic r = 0.82 (p < 0.01), bottled cellular degeneration--r = 0.65 (p < 0.05), lobular inflammation--0.58 (p < 0.05), fibrosis r = 0.67 (p < 0.05), than ALAT--0.51 (p < 0.05), 0.48 (p < 0.05), 0.42 (p < 0.05), 0.51 (p < 0.05) accordingly. Liver venous perfusion worsening was revealed already in moderate SH, when clinical symptoms of portal hypertension were absent. CI in portal vein correlated closely with HAI r = 0.78 (p < 0.05) and fibrosis r = 0.69 (p < 0.05) in NAFLD, that confirmed its diagnostic role in detection of parenchimatous and vascular liver architectonics deterioration and portal hypertension development. CONCLUSION: GGTF is more sensitive parenchimatous damage marker in NAFLD than ALAT and ASAT. Liver venous perfusion study with estimation portal vein congestive index permits to reveal the portal hypertension long before clinical symptoms of this syndrome appearance. Liver parenchimatous damage and its perfusion disturbance confirm the NAFLD progressive course.

[NS3AG expression in chronic hepatitis C: relationship with clinical histological activity and HCV genotypes].

AIM: we have analyzed the peculiarities of NS3Ag expression in hepatocytes and dependence of necroinflam mation on the NS3Ag level and on its localization in hepatocytes in patients with different genotypes of HCV MATERIALS AND METHODS: 31 patients with chronic hepatitis C were examined, 14 (45%) of them with HCV geno type 1b and 17 (55%) with genotype 3a. NS3Ag was detected in the liver tissue by immunohistochemistry ("Novocastra" kit, UK). Routine laboratory analyses were performed. Staging and grading were assessed according to the Knodell score and a modified method, and sclerosis index was determined according to the Metavir scale. RESULTS: the level of NS3Ag in the cytoplasm of hepatocytes was 140 fold higher than in the nucleus. Different patterns of NS3Ag localization in the cytoplasm were observed: in 19 (61%) patients it was detected peripherally near the plasma membrane, whereas in 12 (39%) patients it was diffusely distributed in the cytoplasm. Necroinflammation was more pronounced in the case of peripheral near-membrane NS3Ag localization. The fatty an hydropic degeneration and small cell dysplasia of hepatocytes were more significant in high NS3Ag concentration. No correlation was revealed between the intracytoplasmic NS3Ag localization and the genotype of HCV. However, biochemical activity and the severity of inflammation and fibrosis in all zones of the acinus were highe in patients with HCV genotype 1b, in spite of the lower viral load compared with the patients with genotype 3a CONCLUSIONS: in chronic hepatitis C, cytoplasmatic expression of NS3Ag prevailed over nuclear expression. Sever ity of necroinflammation was higher in patients with peripheral near-membrane NS3Ag localization than in those with uniform diffuse localization. The degree of parenchymatous damage and small cells dysplasia of he patocytes depended on the level of NS3Ag in hepatocytes. In patients with HCV genotype 1b, biochemical an histologic indicators of activity were higher, while NS3Ag concentration was lower compared to genotype 3a.

[Splenoportal blood flow with nonalcoholic fatty liver disease].

UNLABELLED: The aim of this study was to characterise by duplex Doppler sonography the splenoportal venous and arterial blood flow in patients with nonalcoholic fatty liver disease (NAFLD) depending on the grade of biopsy proven steatosis. MATERIALS AND METHODS: 37 patients with NAFLD were examined: 22 female and 15 male average age 46,8 +/- 10,2 (29-62), with IBM 33,5 +/- 4,8 (28-42). The grade of steatosis was estimated by morphological investigation according to Brunt classification (1999). Colour Doppler sonography was performed by the same researcher using Doppler system Vivid-pro-7, USA with a 3,5 MHz convex probe. RESULTS: Alterations were detected in the following parameters: increase of portal and splenic vein diameters, slow-down of blood flow velocity in the portal and splenic veins, increase of the congestion index, decrease of systolic and diastolic blood flow velocity in the common hepatic and splenic artery, increase of pulsatility and resistance indexes of these arteries, increase of portal hypertensive index and decrease of liver vascular index according to the progression of steatosis with maximal negative dynamics of all characteristics in patients with III grade of steatosis. The closest correlative connections were revealed between the grade of steatosis and congestion index (r = 0.81), portal hypertensive index (r = 0.79), and negative connection--with liver vascular index (r = -0.69). CONCLUSION: Using Doppler sonography we have detected a deterioration of venous and arterial splenoportal hemodynamics in patients with nonalcoholic fatty liver disease in connection with the progression of steatosis, therefore this method may be used as a noninvasive way to estimation the grade of steatosis, establish the diagnosis of portal hypertension and detect the formation of liver cirrhosis.

[Diagnostic implications of extracellular matrix proteins in chronic hepatitis and hepatic cirrhosis]. / Diagnosticheskoe znachenie belkov ékstratselliuliarnogo matriksa pri khronicheskom gepatite i tsirroze pecheni.

Enzyme immunoassay measured concentrations of basic extracellular matrix proteins, collagen type 3 (C-3) and fibronectin, in blood plasm of 119 patients with chronic hepatic diseases. 30, 16, 18, 6 and 49 of them had chronic hepatitis of minimal activity (MiACH), that of moderate activity (MACH), intermediate activity (CHIA), high activity (HACH) and hepatic cirrhosis (HC), respectively. The highest C-3 level occurred in C-stage HC, the lowest--in MiACH. Fibronectin was the highest in HACH, minimal--in C-stage HC. C-3 and fibronectin levels correlated with severity of mesenchymal-inflammatory syndrome in CH and HC; in CHIA, HACH and in B-stage HC--with cytolysis markers. A direct relationship was found between protein-synthetizing function of hepatocytes and fibronectin levels in CHIA, HACH and HC while it was inverse in relation to C-3 amount in HC. Thus, tests for plasm C-3 and finronectin expand potentialities of laboratory diagnosis of the process activity in CH and HC, allow prediction of probability of CH transformation into HC.