Cytofluorimetric research on the content of glycogen and its fractions in the hepatocytes of patients with liver cirrhosis of different etiologies]

By cytofluorometric method, a study was made of the total glycogen and its two fractions in liver parenchymal cells both in the donors (20 men) and in patients with cirrhosis of different etiology (39 men). The examination was performed on preparations--smears of isolated hepatocytes, obtained from the live functional liver biopsies. The quantitative analysis has shown an increase in the total glycogen content in hepatocytes of patients with cirrhosis by 3 times compared to the norm, and this increase is independent on the etiology of liver cirrhosis. To study the mechanism of the discovered glycogenosis, the activity of key enzymes of glycogenolyses was determined. It was shown that glucose-6-phosphatase and glycogen-phosphorylase activity in the liver with cirrhosis was lower than in the norm. The most considerable changes were shown in hepatocytes of patients with liver cirrhosis in fractional glycogen composition and, even more significant, in the content of a hard soluble fraction. The hard soluble fraction portion was higher in hepatocytes of the patients with liver cirrhosis of alcohol etiology. The quantitative analysis of glycogen fraction contents in liver cells may be the best marker in the differential diagnosis of symptomless elapsing liver cirrhosis.     

The course of liver cirrhosis not preceded by viral hepatitis with p articular reference to toxic etiology]

The study was aimed at analysing the epidemiological structure of patients with liver cirrhosis without HBsAg treated in 1980-1988. There were 231 of such cases in this period of time. The most frequent cause of liver cirrhosis in patients under 60 years of life was chronic alcoholism whereas 40% of the diagnosed liver cirrhosis in older persons was of unclear etiology. Patients complaints, clinical examinations, and results of the laboratory tests were analysed. The course of the disease was more severe in alcohol-produced liver cirrhosis leading to the haemorrhage from esophageal varices in 36%, and coma in 8% of cases. Alcohol-produced liver cirrhosis promoted other complications such as: cancer of the liver, hepato-renal syndrome or encephalopathy. Liver cirrhosis of unclear etiology in the elderly may be a consequence of the prolonged exposition to environmental pollutants. More severe course of alcohol-produced liver cirrhosis may depend on simultaneous action of two harmful factors: alcohol and environmental pollutants.     

Aging influence on delta-6-desaturase activity and fatty acid composition of rat liver microsomes

The activity of delta-6-desaturase (D6D) in liver microsomes and fatty acid composition of microsomal lipids of rats of different ages were studied. The D6D activity was similar in suckling rats and in weaning rats. However, the enzyme showed a significantly decreased activity in oldest animals, and a linear correlation was found between the D6D activity and the animal age. The fatty acid composition data on total lipids of liver microsomes were consistent with the age-dependent changes in fatty acid desaturase activity. The major changes occurred in the linoleate and arachidonate fractions; the 20:4/18:2 ratio in liver microsomes decreased together with D6D activity during aging. The loss of D6D activity may be a key factor in aging through altering lipid membrane composition.     

Mathematical morphologic analysis of liver cirrhosis. Correlation with etiology, clinical score and hepatocellular carcinoma.

Remodeling of the cirrhotic liver was studied retrospectively by mathematical morphologic methods in 75 autopsy cases (40 alcoholic, 17 hepatitis B virus (HBV)-related and 18 cryptogenetic cirrhosis), including 28 hepatocellular carcinomas. The aim was to obtain objective measurements of cirrhotic patterns that could be correlated with liver function evaluated by the Pugh-Child score, establish the relationship among different morphogenetic features and evaluate the implications of an objective classification of cases by numerical taxonomy in terms of their etiology, liver function and malignant transformation. The results indicate that the Pugh-Child score was closely related to the global amount of fibrosis or to the percentage of regenerative nodules < 0.8 mm in diameter. In contrast, the higher the percentage of lobular-sized regenerative nodules (0.8-1.6 mm), the better the functional score, suggesting that they are probably residual lobules, albeit completely surrounded by fibrous tissue, rather than true regenerative pseudolobules. The four groups of cases obtained by numerical taxonomy (cluster analysis) showed different distributions for alcoholic and HBV-related cirrhosis. The pattern of the latter was practically analogous to that in classically labeled cryptogenetic cirrhosis, suggesting its viral etiology. Taxonomic classification had functional implications. The Pugh-Child score showed a definite relationship with the different clusters obtained. The incidence of malignant transformation gradually decreased from group G1 to G4, with a steeper descent between G2 and G3. These results might contribute to a more dynamic concept of morphologic changes in liver biopsies from patients with cirrhosis.     

Lipidic last breath of life in patients with alcoholic liver disease

Alcoholic liver disease (ALD) begins with the accumulation of lipid droplets in the liver. Lipids which accumulate in the liver can stimulate inflammation, and the fatty acid derivatives, hydroxyeicosatetraenoic acids (HETEs) and hydroxyoctadecadienoic acids (HODEs), may play an important role in this process. We evaluated the concentrations of linoleic and arachidonic acid derivatives in the plasma of patients with ALD, non-alcoholic fatty liver disease (NAFLD) and healthy individuals. The groups consisted of 173 subjects: 63 patients with ALD, 90 with NAFLD and 20 healthy volunteers. Plasma 12-, 15-, and 5-HETE as well as 9- and 13-HODE were assessed using HPLC and isoprostane 8-epi-PGF 2α III was evaluated with an ELISA. In addition the mRNA expression of lipoxygenases (5-LOX, 15-LOX-1, 15-LOX-2) in the liver samples of patients with ALD cirrhosis was measured. A significant difference between the plasma concentrations of the analyzed derivatives was found when divided according to gender. The most significant differences were found between healthy individuals and ALD patients, as well as ALD and NAFLD individuals regardless of gender. The increased plasma HODEs and HETEs concentrations were in line with the increase in 5- and 15-LOX-1 and 15-LOX-2 mRNA in liver samples from ALD cirrhosis patients. LOXs expression and peroxidation of polyunsaturated fatty acids by free radical-propagated chemical oxidation may be contributing factors in liver necroinflammatory injury in ALD.     

不同病因肝硬化患者L3骨骼肌指数特征及其对患者营养状况的预测价值分析

摘要 目的：探讨不同病因肝硬化患者L3骨骼肌指数特征及其对患者营养状况的预测价值分析。方法：选取2019年6月-2022年6月在我院收治的120例肝硬化患者作为研究对象,其中乙肝肝硬化40例,酒精性肝硬化40例,自身免疫性肝炎肝硬化40例。比较乙肝肝硬化组,酒精性肝硬化组,自身免疫性肝炎肝硬化L3 SMI的特征。采用Pearson相关检验分析L3 SMI与肝硬化患者营养状况的相关性。采用Logistics回归模型构建影响肝硬化营养状况的独立危险因素。采用受试者工作曲线（ROC）评估L3 SMI对肝硬化营养状况的预测价值。结果：自身免疫性肝炎肝硬化组L3 SMI、25（OH）D、ALB、PA、TRF的表达水平均显著低于酒精性肝硬化组和乙肝肝硬化组（P<0.05）,且酒精性肝硬化组显著低于乙肝肝硬化组（P<0.05）。肝硬化患者LSM与25（OH）D、ALB、PA、TRF均显著正相关（P<0.05）。以肝硬化患者营养状况作为因变量（营养正常=0,营养不良=1）纳入logistics回归模型,结果显示,25（OH）DALB、PA、TRF、L3 SMI是危险因素（P<0.05）。多因素分析结果显示,25（OH）DALB、PA、TRF、L3 SMI是影响肝硬化患者营养状况的独立危险因素（P<0.05）。L3 SMI预测评估肝硬化患者营养状况的Youden指数0.765,敏感度85.00（%）,特异度82.00（%）,AUC值0.810,95%CI：0.685~0.912。结论：不同病因肝硬化患者L3 SMI存在明显差异,临床可采用L3 SMI对肝硬化患者营养状况做出预测评估。     

Atrial natriuretic peptide-renin-aldosterone system in cirrhosis of the liver: circadian study

Plasma levels of immunoreactive atrial natriuretic peptide (ANP), plasma renin activity (PRA), and plasma aldosterone (PA) were measured for an entire day at 6:00 am, 8:00 am, 12:00 pm, 6:00 pm, 8:00 pm, and 12:00 am in 6 healthy subjects, in 10 patients with compensated cirrhosis of the liver, and in 10 cirrhotics with ascites. The subjects, after synchronized standard life conditions lasting for 6 days were held in a clinostatic position during the study. The data were analyzed by the "cosinor" method. The results show significant circadian rhythms for the three biological variables in healthy subjects. In the compensated cirrhotic group, a circadian rhythm was detected only for PA. No rhythm was demonstrated in the ascitic patients. These data suggest that in cirrhosis of the liver, great variations in secretion rhythmicity for PRA and ANP are present, while maintaining the intrinsic PA rhythmicity, which is lost in patients with ascites. This progressive derangement in PA circadian rhythm in the ANP-PRA-PA system can be considered as an index of evolution in the natural history of cirrhosis of the liver.     

In vitro formation of fibrous septa by liver connective tissue cells

Summary Active fibrous septa are a common feature in liver fibrosis and cirrhosis. Their etiology and formation were studied using cultures of tissue fragments or cells included in collagen gels. Liver fragments obtained from patients with cirrhosis or severe schistosomal fibrosis were able to reorganize the gel and to form discrete, interconnecting fibrous septa composed of parallel arrays of collagen, subsequently colonized by migrating connective tissue cells. The same was obtained in cultures of fibrogranulomatous lesions isolated from schistosome-infected mice livers. However, fragments of normal human and murine liver tissue did not show the capacity to form fibrous septa. Septa formation was also obtained in cultures of cell spheroids formed by liver connective tissue cells isolated from human fibrotic or cirrhotic liver tissues, but not with spheroids of normal skin fibroblasts or smooth muscle cells. This experimental model may represent the fibrous septa formation in vivo, depending on the activity of liver connective tissue cells. The ability of tissue fragments or cell spheroids to form septa in collagen gels might reflect the degree of fibrosis present in the liver tissue in vivo. This research was supported by FINEP and CNPq (Brazil) and CNRS (France).     

Serum alpha-mannosidase in patients with alcoholic liver disease

Sera from 9 persons with either biopsy-proven alcoholic liver disease or a history of chronic, excessive ethanol consumption were analyzed for their content of various hydrolases. Compared to controls, significant elevations in the following enzyme activities were seen in sera from the patient population: acid phosphatase (2.0-fold), beta-glucuronidase (2.1-fold), hexosaminidase (1.4-fold), and alpha-L-fucosidase (2.3-fold). In addition, alpha-mannosidase activity, previously reported to be unchanged in cases of hepatic cirrhosis [Reglero et al., Clinica chim. Acta 130: 155-158], (1980) was found to be significantly increased (p less than 0.001) when assays were performed at acid (pH 4.5) or intermediate (pH 5.5) hydrogen ion concentrations. Fractionation of sera on DEAE-Sephadex columns showed that the increase in alpha-mannosidase activity in the serum of patients with alcoholic liver disease was due to increases in the level of at least one 'acid alpha-mannosidase' and two intermediate pH optimum alpha-mannosidases. The general increase in the activity of a group of glycosidases is consistent with a hypothesis involving decreased clearance of glycoproteins from the blood of persons with hepatic cirrhosis.     

Collagenase of hepatocytes and sinusoidal liver cells in the reversibility of experimental cirrhosis of the liver

In order to explore the cellular source(s) and the behaviour of the collagenolytic activity previously described in rat liver homogenates, in the reversibility of experimental cirrhosis of the liver, enriched suspensions of hepatocytes and of sinusoidal liver cells were obtained by a procedure which employs low EDTA concentrations and no bacterial collagenase. Cell suspensions were prepared from three different groups of animals: 1) normal controls, 2) rats with CCl4-induced cirrhosis of the liver, and 3) rats with swine serum-induced cirrhosis of the liver. Animals were sacrificed in each group upon completion of treatment and also after 3, 6 and 12 months. In each liver wet weight and collagen concentration were determined, and collagenolytic activity of both enriched cell suspensions was measured separately. In addition, histological studies of liver tissue and ultrastructural examination of cell suspensions were performed by standard procedures. Enriched suspensions of both normal hepatocytes and sinusoidal liver cells display Ca2(+)-dependent collagenolytic activities. Both cell suspensions obtained from each of the two types of cirrhotic livers show normal or slightly increased average levels of collagenase activity at the time of treatment discontinuation, when average liver collagen content ranges from 6 to 10-fold over normal, suggesting that the normal collagenase/collagen ratio is disturbed and that collagenolytic activity is deeply decreased in relation to the actual liver collagen load.(ABSTRACT TRUNCATED AT 250 WORDS)     

Correlation of endothelin-1 concentration and angiotensin-converting enzyme activity with the staging of liver fibrosis

Increased serum angiotensin-converting enzyme (SACE) activity and serum concentration of endothelin-1 (ET-1) were found in liver cirrhosis. We investigated a correlation between the different stages of liver fibrosis and SACE activity and serum ET-1 concentration. Seventy patients with pathohistologically established chronic liver disease were divided in three groups according to Ishak criteria for liver fibrosis: minimal fibrosis (Ishak score 0-1, n =20), medium fibrosis (Ishak score 2-5, n=20) and cirrhosis (Ishak score 6, n=30). SACE activity and ET-1 concentration were determined using commercial ELISA kits. SACE activity and ET-1 concentrations were proportional to the severity of disease, the highest being in patients with liver cirrhosis. Maximal increase in SACE activity was found between minimal and medium fibrosis while maximal increase in ET-1 concentration was revealed between medium fibrosis and cirrhosis. The analysis of the Receiver Operating Characteristic (ROC) curve for SACE activity suggested a cut-off value to separate minimal from medium fibrosis at 59.00 U/L (sensitivity 100%, specificity 64.7%). The cut-off value for serum ET-1 concentration to separate medium fibrosis from cirrhosis was 12.4 pg/mL (sensitivity 96.8%, specificity 94.4%). A positive correlation between SACE activity and ET-1 concentration was registered (Spearman's ? = 0.438, p = 0.004). Both SACE activity and ET-1 concentration were increased in all stages of liver fibrosis. Cut-off points for SACE activity and ET-1 concentration could be a biochemical marker for the progression of fibrosis. Positive correlation between SACE activity and ET-1 concentration might indicate their interaction in the development of liver cirrhosis.     

Impaired hepatic removal of interleukin-6 in patients with liver cirrhosis

Systemic concentrations of interleukin-6 (IL-6) are elevated in patients with liver cirrhosis, and impaired hepatic uptake of IL-6 was suggested to contribute to higher levels in these patients. To test this hypothesis IL-6 was measured in portal venous serum (PVS), hepatic venous serum (HVS) and systemic venous serum (SVS) of 41 patients with liver cirrhosis and four patients with normal liver function. IL-6 was higher in PVS than HVS of all blood donors and about 43% of portal vein derived IL-6 was extracted by the healthy liver, and 6.3% by the cirrhotic liver demonstrating markedly impaired removal of IL-6 by the latter. Whereas in patients with CHILD-PUGH stage A IL-6 in HVS was almost 25% lower than in PVS, in patients with CHILD-PUGH stage C IL-6 was similarly abundant in the two blood compartments. Ascites is a common complication in cirrhotic patients and was associated with higher IL-6 levels in all blood compartments without significant differences in hepatic excretion. Hepatic venous pressure gradient did not correlate with the degree of hepatic IL-6 removal excluding hepatic shunting as the principal cause of impaired IL-6 uptake. Furthermore, patients with alcoholic liver cirrhosis had higher IL-6 in all blood compartments than patients with cryptogenic liver cirrhosis. Aetiology of liver cirrhosis did not affect hepatic removal rate indicating higher IL-6 synthesis in patients with alcoholic liver cirrhosis. In summary, the current data provide evidence that impaired hepatic removal of IL-6 is explained by hepatic shunting and liver dysfunction in patients with liver cirrhosis partly explaining higher systemic levels.     

Cytidine-5'-monophosphate-N-acetylneuraminic acid. Asialoglycoprotein sialic acid transferase activity in liver and serum of patients with juvenile hepatic cirrhosis and alpha-1-antitrypsin deficiency.

The molecular basis for the accumulation of a substance which displays the immunological reactivity of alpha-1-antitrypsin within vesicles of liver parenchymal cells of individuals with hepatic cirrhosis and serum alpha-1-antitrypsin deficiency remains unclear. We recently reported that serum from a patient with alpha-1-antitrypsin deficiency and hepatic cirrhosis was substantially deficient in sialyltransferease (EC 2.4.99.1) an enzyme which transfers sialic acid from cytidine 5'-monophosphate-N-acetylneuraminic acid to a variety of asialoglycoprotein acceptors. In the present report we have extended these studies to include serum from five additional patients with alpha-1-antitrypsin deficiency and juvenile hepatic cirrhosis as well as a liver specimen obtained at autopsy of one of these patients. We find the sialytransferase activity in serum from six patients with alpha-1-antitrypsin deficiency and hepatic cirrhosis to be 50% of healthy pediatric control values and 30% of pediatric patients with liver disease. However, serum from family members homozygous for alpha-1-antitrypsin deficiency but without hepatic cirrhosis, and serum from patients with a variety of other kinds of liver disease, failed to exhibit the marked sialytransferase deficiency. Similar assays carried out on a homogenate of a liver sample from one patient with alpha-1-antitrypsin deficiency and hepatic cirrhosis indicated that the deficiency of sialyltransferase activity was not demonstrable in liver. Furthermore, a comparative kinetic analysis of serum and liver sialytransferase in normal and afflicted individuals failed to detect differences in substrate affinities which might account for a decrease in functional sialyltransferase capacity in individuals with alpha-1-antitrypsin deficiency and hepatic cirrhosis. These observations suggest that the serum sialyltransferase deficiency in such patients probably arises after chronic and extensive liver disease involving hepatic accumulation of alpha-1-antitrypsin rather than the enzyme deficiency being the primary cause of the hepatic cirrhosis and alpha-1-antitrypsin deficiency.     

Decreased serum osteocalcin levels in patients with liver cirrhosis

Serum levels of osteocalcin (OC) have been found to be a specific biochemical parameter of bone formation. We measured serum levels of osteocalcin, parathyroid hormone (PTH) and 25-hydroxyvitamin D (25(OH)D) in 49 patients with liver cirrhosis, who are known to have an increased prevalence of metabolic bone disease, and a matched control group (n = 35). Serum levels of OC were significantly decreased in the patients with liver cirrhosis when compared to control subjects (P < 0.001). Serum levels of 25(OH)D were decreased (P < 0.001), whereas no statistical difference was found between the serum levels of PTH in the patients with liver cirrhosis and those of the controls. In a subgroup of 23 patients with cirrhosis of the liver and 34 control subjects, the bone mineral content (BMC) of the non-dominant forearm was determined by single photon absorptiometry. BMC was significantly lower in the patient with liver cirrhosis than the control subjects (P < 0.04). Our data demonstrate vitamin D deficiency, decreased bone formation and a decreased BMC in patients with liver cirrhosis.     

Plasma clearance of cholic acid in patients with chronic diseases of the liver

The diagnostic value of 14C-cholic acid plasma clearance following oral administration was evaluated. 14C-cholic acid clearance was 1223 +/- 267 mL/min. per 1 square meter in the control group without liver disease. Significantly lower values (p less than 0.001) were obtained in the patients with chronic hepatitis (694 +/- 137 mL/min. per 1 square meter) and liver cirrhosis (332 +/- 156 mL/min. per 1 square meter). Sensitivity of the 14C-cholic acid clearance test was 100% while specificity--80%. A 3-year follow-up has shown that this test is of high prognostic value in patients with liver cirrhosis.     

Extracellular breakdown of collagen as affected by lysosomal enzymes during the involution of liver cirrhosis

Electron microscopy and electron histochemistry (exposure to acid phosphatase) were used to study the mechanisms of extracellular degradation of collagen in the liver during involution of experimental cirrhosis. The following results were obtained: extracellular secretion of lysosomal enzymes from hepatocytes and connective tissue cells takes place in liver cirrhosis and its involution; partial hepatectomy during liver cirrhosis stimulates the activity of acid phosphatase in the liver cells; the lysosomal enzymes, excreted from hepatocytes and connective tissue cells by means of exocytosis take an active part in collagen extracellular degradation in vivo; at initial stages of cirrhosis involution extracellular degradation of collagen in the liver occurs at the expense of lysosomal enzymes from hepatocytes and connective tissue cells. Subsequently, as cirrhosis regresses, the principal role in the lysis of collagen gradually passes to lysosomal enzymes of hepatocytes.     

Amino acid composition of human liver mitochondrial membranes in normal and pathological conditions

The amino acid composition of proteins from liver mitochondrial membranes has been studied in patients with normal liver, with biliary diseases and fatty liver, with obstructive jaundice or liver cirrhosis. A characteristic pattern of the amino acid composition in patients with normal liver has been found. In the mitochondrial membranes of patients with fatty liver tryptophan and lysine were decreased while [aspartic acid plus asparagine] and [glutamic acid plus glutamine] were increased compared to their counterpart in the normal liver. In patients with obstructive jaundice of short duration (less than two months) only a slight decrease in methionine content was found, while in the case of liver cirrhosis amino acid composition was markedly changed.deceased.     

Age-related changes in linoleate and alpha-linolenate desaturation by rat liver microsomes

The first and rate limiting step in the conversion of linoleic and alpha-linolenic acid is catalyzed by the delta - 6 - desaturase (D6D) enzyme. Rat liver microsomal D6D activity decreases on linolenic acid at a rate proportional to the animal age; on alpha-linolenic acid the decrease in D6D activity begins only later than on linoleic acid. The fatty acid composition of liver microsomes determined by gas chromatographic analysis confirms the impairment of the enzymatic activity directly measured. Our data indicate a correlation between aging and D6D activity impairment. The loss of D6D activity may be a key factor in aging through altering the eicosanoid balance.     

Serum haptoglobin type and liver cirrhosis.

Haptoglobin phenotypes were studied by a polyacrylamide gel electrophoresis on 200 blood donors and 105 patients with liver cirrhosis, of which 79% belonged to non-alcoholic etiology. Though no difference of haptoglobin types could be found between blood donors with positive and negative hepatitis B antigen, the cirrhois patients had an excess haptoglobin gene 1. The patients with haptoglobin gene 1 were associated with severe liver dysfunction. Since the family pedigrees of the patients with type 1--1 excluded individuals with type 2--2, the phenotypes seemed to be stable in the cirrhotic process. The possibility that the haptoglobin 2 gene offered resistence to the non-alcoholic cirrhosis was discussed.     

Acid phosphatase distribution in the liver in cirrhosis

The distribution of acid phosphatase in liver cirrhosis, as well as in its reverse development, was investigated in mice using histochemistry and electron histochemistry methods. Histochemistry demonstrated a sharp activity increase of acid phosphatase (as compared with the same in the material of partial hepatectomy) in liver cells (especially hepatocytes) during liver cirrhosis regression 10 days after a partial hepatectomy. Electron histochemistry has shown the enzyme withdraw out of hepatocytes and connective tissue cells of fibrotic stratum in the extra-cell medium. The reaction product localized on the neighbouring collagen fibres giving evidence that during reverse development of liver cirrhosis the lisosomal enzyme release from specified cells by means of exocytosis and they are involved in the lysis of collagen.