Network pharmacology assessment of Qingkailing injection treatment of cholestatic hepatitis.

OBJECTIVE: To investigate the targets and mechanisms of action of Qingkailing injection (，QKL) in the treatment of cholestatic hepatitis. METHODS: A network pharmacology method was implemented using drug and disease databases to target QKL and cholestasis hepatitis, respectively. The functional protein association network STRING database was used to construct a protein-protein interaction network using R language and the Bioconductor toolkit. The org.Hs.eg.db and clusterProfiler packages were used for gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, which explored biological functions and pathways of potential targets. Targets were then visualized using Cytoscape 3.6.0 software. RESULTS: We screened 121 compounds in QKL and identified 112 targets for the treatment of cholestatic hepatitis. QKL played a role in the treatment of cholestatic hepatitis through 305 biology process terms, 15 cellular component and 29 molecular function terms. The mechanism of QKL action was mainly related to tumor necrosis factor, mitogen-activated protein kinase, and PI3K-Akt signaling pathways. CONCLUSION: The treatment of cholestatic hepatitis by QKL involved multiple targets, biological functions, and signaling pathways that are closely associated with the disease.

Assessment of Adenosine Triphosphatase Phospholipid Transporting 8B1 (ATP8B1) Function in Patients With Cholestasis With ATP8B1 Deficiency by Using Peripheral Blood Monocyte-Derived Macrophages.

Adenosine triphosphatase phospholipid transporting 8B1 (ATP8B1) deficiency, an ultrarare autosomal recessive liver disease, includes severe and mild clinical forms, referred to as progressive familial intrahepatic cholestasis type 1 (PFIC1) and benign recurrent intrahepatic cholestasis type 1 (BRIC1), respectively. There is currently no practical method for determining PFIC1 or BRIC1 at an early disease course phase. Herein, we assessed the feasibility of developing a diagnostic method for PFIC1 and BRIC1. A nationwide Japanese survey conducted since 2015 identified 25 patients with cholestasis with ATP8B1 mutations, 15 of whom agreed to participate in the study. Patients were divided for analysis into PFIC1 (n = 10) or BRIC1 (n = 5) based on their disease course. An in vitro mutagenesis assay to evaluate pathogenicity of ATP8B1 mutations suggested that residual ATP8B1 function in the patients could be used to identify clinical course. To assess their ATP8B1 function more simply, human peripheral blood monocyte-derived macrophages (HMDMs) were prepared from each patient and elicited into a subset of alternatively activated macrophages (M2c) by interleukin-10 (IL-10). This was based on our previous finding that ATP8B1 contributes to polarization of HMDMs into M2c. Flow cytometric analysis showed that expression of M2c-related surface markers cluster of differentiation (CD)14 and CD163 were 2.3-fold and 2.1-fold lower (95% confidence interval, 2.0-2.5 for CD14 and 1.7-2.4 for CD163), respectively, in patients with IL-10-treated HMDMs from PFIC1 compared with BRIC1. Conclusion: CD14 and CD163 expression levels in IL-10-treated HMDMs may facilitate diagnosis of PFIC1 or BRIC1 in patients with ATP8B1 deficiency.

Assessment of Selected Parameters of Liver Fibrosis and Inflammation in Patients with Diagnosed Cystic Fibrosis.

Changes in the liver and bile ducts observed in patients diagnosed with cystic fibrosis result from inflammatory processes as well as fibrosis, remodeling, apoptosis, and cholestasis. As a consequence, portal hypertension, cirrhosis, and hepatic failure may develop. So far, the complexity of these processes has not been elucidated. Study Objectives. The aim of the study was to evaluate the selected parameters of hepatitis and fibrosis (Fibrotest, Actitest, and APRI) in patients diagnosed with cystic fibrosis. Material and Methods. The study included 79 patients with cystic fibrosis, aged 1 to 20 years (mean age 9.8 years), 49 girls (62%) and 30 boys (38%). The analysis involved the following: age, sex, clinical manifestations, laboratory tests evaluating pancreas function, parameters of liver damage, and cholestasis. Fibrotest, Actitest, and APRI were performed in all subjects. Results. Elevated parameters of hepatic cell damage (hypertransaminasemia) were found in 31/79 (39.2%) patients, while abnormal cholestasis parameters in 21/79 (26.6%). The abnormal results of Fibrotest were reported in 15% of patients (12/79), while of Actitest in 10% (8/79). In contrast, elevated APRI values were found in only 7.6% (6/79) of subjects. There was a statistically significant correlation between APRI and age (higher values were observed in younger children) and between Fibrotest and Actitest and pancreatic insufficiency (higher values were found in subjects without this abnormality). Moreover, Fibrotest values were significantly higher in girls. There was no correlation between Fibrotest, Actitest, and APRI values and the type of mutation. Conclusion. It appears that Fibrotest may be used as an early marker of liver fibrosis in patients with cystic fibrosis. Increased APRI values were only found in subjects with advanced hepatic lesions, most often in the form of portal hypertension.

Organotypic 3D HepaRG Liver Model for Assessment of Drug-Induced Cholestasis.

Cholestasis remains a major challenge in drug-induced liver injury, and therefore warrants identification of chemical entities that may lead to cholestasis. Recent advances in cell culture methods enable 3D spheroid models to remain viable for much longer periods of time than conventional sandwich cultures of primary human hepatocytes while maintaining native tissue-like functionality, such as drug metabolism activity, receptor signaling functionality, and physiological relevance. These spheroid models enable us to study repeated exposure effects associated with chemicals and their metabolites that may ultimately progress to cholestasis and liver injury. HepaRG cells cultured as spheroids are viable for more than 4 weeks with cytochrome P450 enzymatic activities comparable to ranges observed in freshly isolated/cryopreserved suspensions of primary human hepatocytes. HepaRG spheroids form bile canalicular structures with potential application as a model to study biliary excretion processes and intrahepatic obstruction of bile flow, leading to hepatocellular damage and death. In this chapter, we describe methods to culture 3D spheroids of HepaRG cells with extensive bile canalicular structures/networks, image transport of bile acid (cholyl-lysyl-fluorescein) to the bile canaliculi, and measure cholestatic drug-induced cytotoxicity.

Serum and Hepatic Autofluorescence as a Real-Time Diagnostic Tool for Early Cholestasis Assessment.

While it is well established that various factors can impair the production and flow of bile and lead to cholestatic disease in hepatic and extrahepatic sites, an enhanced assessment of the biomarkers of the underlying pathophysiological mechanisms is still needed to improve early diagnosis and therapeutic strategies. Hence, we investigated fluorescing endogenous biomolecules as possible intrinsic biomarkers of molecular and cellular changes in cholestasis. Spectroscopic autofluorescence (AF) analysis was performed using a fiber optic probe (366 nm excitation), under living conditions and in serum, on the livers of male Wistar rats submitted to bile duct ligation (BDL, 24, 48, and 72 h). Biomarkers of liver injury were assayed biochemically. In the serum, AF analysis distinctly detected increased bilirubin at 24 h BDL. A continuous, significant increase in red-fluorescing porphyrin derivatives indicated the subversion of heme metabolism, consistent with an almost twofold increase in the serum iron at 72 h BDL. In the liver, changes in the AF of NAD(P)H and flavins, as well as lipopigments, indicated the impairment of mitochondrial functionality, oxidative stress, and the accumulation of oxidative products. A serum/hepatic AF profile can be thus proposed as a supportive diagnostic tool for the in situ, real-time study of bio-metabolic alterations in bile duct ligation (BDL) in experimental hepatology, with the potential to eventually translate to clinical diagnosis.

Assessment of ATP8B1 Deficiency in Pediatric Patients With Cholestasis Using Peripheral Blood Monocyte-Derived Macrophages.

Progressive familial intrahepatic cholestasis type 1 (PFIC1), a rare inherited recessive disease resulting from a genetic deficiency in ATP8B1, progresses to liver failure. Because of the difficulty of discriminating PFIC1 from other subtypes of PFIC based on its clinical and histological features and genome sequencing, an alternative method for diagnosing PFIC1 is desirable. Herein, we analyzed human peripheral blood monocyte-derived macrophages (HMDM) and found predominant expression of ATP8B1 in interleukin-10 (IL-10)-induced M2c, a subset of alternatively activated macrophages. SiRNA-mediated depletion of ATP8B1 in IL-10-treated HMDM markedly suppressed the expression of M2c-related surface markers and increased the side scatter (SSC) of M2c, likely via impairment of the IL-10/STAT3 signal transduction pathway. These phenotypic features were confirmed in IL-10-treated HMDM from four PFIC1 patients with disease-causing mutations in both alleles, but not in those from four patients with other subtypes of PFIC. This method identified three PFIC1 patients in a group of PFIC patients undiagnosed by genome sequencing, an identical diagnostic outcome to that achieved by analysis of liver specimens and in vitro mutagenesis studies. In conclusion, ATP8B1 deficiency caused incomplete polarization of HMDM into M2c. Phenotypic analysis of M2c helps to identify PFIC1 patients with no apparent disease-causing mutations in ATP8B1.

Critical Factors in the Assessment of Cholestatic Liver Injury In Vitro.

Cholestasis is a common pathological component of numerous liver diseases. The initiating event during cholestatic liver injury is widely believed to be the accumulation of bile acids in hepatocytes and the hepatic parenchyma. As bile acids are considered the primary toxic compounds in the injury, numerous in vitro models of bile acid-induced injury and bile acid-induced changes in gene expression have been developed to attempt to better define cholestasis at a cellular level. This chapter focuses on the establishment of a system for determining the effects of cholestatic concentrations of bile acids on hepatocytes using primary hepatocytes or hepatoma cell lines. Moreover, this chapter addresses significant differences in the response of different species to bile acid exposure and novel information on the relevance of treating hepatocytes with concentrations of specific bile acids.

Improved cardiac function in patients with obstructive jaundice after internal biliary drainage: hemodynamic and hormonal assessment.

OBJECTIVE: To investigate myocardial function in patients with obstructive jaundice before and after internal biliary drainage. SUMMARY BACKGROUND DATA: Increased plasma levels of atrial natriuretic peptide (ANP) have been found in patients with biliary obstruction. METHODS: Thirteen patients with newly diagnosed obstructive jaundice and no previous heart, lung, or renal disease were studied using a Swan-Ganz catheter. Hemodynamic measurements were taken before and 4 days after internal biliary drainage. Levels of ANP and brain natriuretic peptide (BNP) were obtained and liver function tests were also determined. RESULTS: Plasma levels of ANP and BNP were increased twofold to fourfold in the basal state and declined after biliary drainage. Independent variables predicting left ventricular systolic work were total bilirubin concentrations, duration of jaundice, and BNP. In addition, bilirubin concentrations correlated with pulmonary vascular resistance, mean arterial pulmonary pressure, and right ventricular systolic work. Internal biliary drainage resulted in an improvement in left ventricular systolic work. A correlation was found between decreasing ANP concentrations and increasing cardiac output. CONCLUSIONS: Increased plasma levels of natriuretic peptides in patients with obstructive jaundice may reflect a subclinical myocardial dysfunction correlating with the degree of jaundice. After internal biliary drainage, there is a measurable improvement of cardiac function.

Renal effects of experimental obstructive jaundice: morphological and functional assessment.

BACKGROUND: The pathophysiology of renal impairments occurring in obstructive jaundice has been extensively studied, but the underlying mechanism of these derangements remains unclear. The aim of the present study was to investigate the time-related morphological and functional changes occurring in the kidneys of rats undergoing obstructive jaundice. METHODS: Histological examination, renal function assessment and determination of (Na + K)-ATPase activity were performed in the kidneys of rats 7, 14, and 21 days following bile duct ligation (BDL) or sham operation (sham). RESULTS: Glomerular filtration rate was unaffected by BDL throughout the period of the study. Tubular effects occurred at days 7 and 14, being more marked at day 7, and consisted of an increase of about twice in the fractional excretion of sodium and chloride, paralleled by a decreased proximal and distal tubular reabsorption of sodium of about 50 and 40%, respectively. Natriuresis was consistent with augmentation of osmolar clearance but it was not associated with changes in the activity of renal (Na+ + K+)-ATPase. The ability to dilute urine was impaired at days 14 and 21 after BDL. Additionally, these effects were accompanied by decreased tubulointerstitial fibrosis and vasodilation of inner medullary capillaries. At day 21, the parameters of tubular function in BDL and sham groups were not significantly different. CONCLUSIONS: These data support the view that raised natriuresis taking place in the initial 2 weeks following BDL is due mainly to tubular effects. The contribution of hemodynamic, paracrine and humoral mediators is discussed.

Renal effects of experimental obstructive jaundice: morphological and functional assessment

Background. The pathophysiology of renal impairments occurring in obstructive jaundice has been extensively studied, but underlying mechanism of these derangements remains unclear. The aim of the present study was to investigate the time-related morphological and functional changes occurring in the kidneys of rats undergoing obstructive jaundice. Methods. Histological examination, renal function assessment and determination of (Na + K)-ATPase activity were performed in the kidneys of rats 7, 14, and 21 days following bile duct ligation (BDL) or sham operation (sham). Results. Glomerular filtration rate was unaffected by BDL throughout the period of the study. Tubular effects occurred at days 7 ant 14, being more marked at day 7, and consisted of an increase of about twice in the fractional excretion of sodium and chloride, paralleled by a decreased proximal and distal tubular reabsorption of sodium of about 50 and 40 percent, respectively. Natriuresis was consistent with augmentation of osmolar clearance but it was not associated with changes in the acivity of renal (Na+ + K+)-ATPase. The ability to dilute urine was imparied at days 14 and 21 after BDL. Additionally, these effects were accompanied by decreased tubulointerstitial fibrosis and vasodilation of inner medullary capillaries. At day 21, the parameters of tubular function in BDL and sham groups were not significantly different. Conclusions. These data support the view that rasied natriuresis taking place in the initial 2 weeks following BDL is due mainly to tubular effects. The contribution of hemodynamic, paracrine and humoral mediators is discussed

Preoperative assessment of body fluid disturbances in patients with obstructive jaundice.

Postoperative renal dysfunction in obstructive jaundice (OJ) patients has been associated with hypovolemia and depletion of the extracellular water compartment (ECW). The aim of the study was to evaluate the preoperative status of body compartments in OJ patients measured by two methods. In a prospective study 39 OJ patients (11 benign and 28 malignant obstructions) were investigated, with 15 healthy subjects used as a control group (CG). Bioelectrical impedance analysis (BIA) determinations and values derived from anthropometric measurements were used to assess body compartment status. The coefficient of variation of BIA was below 4% in both OJ and CG subjects. No differences were found in intracellular water. However total body water (TBW) and ECW were reduced in OJ patients (50.5 +/- 4.6 vs. 56 +/- 8% body weight, p = 0.05; and 21 +/- 4.5 vs. 23.8 +/- 2.5% body weight, p < 0.05, respectively). There were no differences between benign and malignant obstructions. Seventy four percent of OJ patients had an ECW volume below the mean +/- 2 SD in the CG subjects. Anthropometric and BIA determinations correlated closely for TBW measurements in both CG (r = 0.92, p < 0.001) and OJ patients (r = 0.91, p < 0.001). Bland-Altman analysis also showed that for TBW the BIA was in agreement with anthropometry. In the present study, BIA offered a good correlation with anthropometric determinations and was a reliable method for body fluid disturbances assessment in jaundiced patients.

Assessment of energy status and fructose metabolism of liver in obstructive jaundice by 31P magnetic resonance spectroscopy.

The effect of cholestasis on hepatic energy status and fructose metabolism in jaundiced rats and patients was investigated using 31P magnetic resonance spectroscopy. Rats with obstructive jaundice (OJ group) were studied 7 days after bile duct ligation. Drainage rats were studied at 3 days (DR3 group) and 7 days (DR7 group) after the relief of 1 week obstruction of the common bile duct. In the bile duct ligated rat, the beta-adenosine triphosphate (ATP)/Pi (inorganic phosphate) ratio was significantly lower than in sham-operated controls. This ratio recovered rapidly in the DR3 and DR7 groups. The maximum increase in the phosphomonoester peak (PMEmax) after an intravenous bolus of fructose was significantly reduced in both the OJ and DR3 groups, and was accompanied by a decrease in hepatic fructokinase activity. The PMEmax and the fructokinase activity recovered in the DR7 group. In a clinical study, the beta-ATP/Pi ratio in six healthy volunteers was comparable to that of 15 patients with obstructive jaundice, regardless of their biliary drainage status. The PMEmax in all patients (serum bilirubin > or = 5 mg/dL), irrespective of biliary drainage, was significantly lower than in volunteers. Furthermore, the PMEmax in four of the eight patients with biliary drainage (serum bilirubin < 5 mg/dL) was lower than in volunteers. It is concluded that while energy status in jaundiced patients is well maintained, fructose phosphorylation is inhibited and recovery is delayed after the relief of obstruction compared with serum bilirubin. For the non-invasive evaluation of damaged liver function in jaundice, 31P magnetic resonance spectroscopy is a useful technique.

Hepatic functional reserve in patients with obstructive jaundice: an assessment by the redox tolerance test.

PURPOSE: Hepatic mitochondrial response to oral glucose load (redox tolerance test) was evaluated as an indicator of hepatic functional reserve of patients with obstructive jaundice. PATIENTS AND METHODS: The redox tolerance test was performed in 29 patients with obstructive jaundice before percutaneous transhepatic biliary drainage and 2 weeks after the procedure. RESULTS: The redox tolerance index (RTI) before drainage was not related to conventional parameters other than albumin, but was significantly associated with bilirubin half-life (P < 0.01). Of 19 patients with an RTI > or = 0.5 before drainage, all maintained similar values after drainage and experienced satisfactory clinical courses, even after major surgery. Of 10 patients with an RTI < 0.5 before drainage, 5 showed improvement and 5 deteriorated after drainage. Four of the latter 5 died within 60 days after drainage. The hospital mortality was significantly greater in patients with initial RTI < 0.5 than in patients with RTI > or = 0.5 (P < 0.01). CONCLUSION: The redox tolerance test is useful for evaluating hepatic functional reserve and prognosis in patients with obstructive jaundice.

Kinetic analysis of impaired work-cost performance in jaundiced rabbit liver.

Impairment of energy metabolism was studied in jaundiced rabbit liver by kinetic analysis of energy transfer function. Free cytosolic ADP (ADPf), as calculated from the measured components of the glyceraldehyde-3-phosphate dehydrogenase and 3-phosphoglycerate kinase/lactate dehydrogenase reactions, decreased from the control value of 48.1 to 37.0 microM at 24 h after bile duct ligation. The maximal velocity (Vmax) of ATP synthesis, as measured by state 3 respiration of isolated mitochondria, decreased from the control value of 62.1 to 38.3 nmol ATP synthesized per min per mg mitochondrial protein, while the Michaelis constant for ADP (Km) decreased from the control value of 19.2 to 12.8 microM. ATP synthesis velocity in vivo }v: Vmax/[1 + (Km/[ADPf])], as calculated by Vmax, Km and ADPf, decreased from the control value of 44.4 to 28.5 nmol ATP synthesized per min per mg mitochondrial protein. Delta v/delta ADPf(delta v/delta ADPf: Vmax.Km/(Km + [ADPf])2), which indicates work-cost performance of the liver, decreased from the control value of 0.263 to 0.198. Biochemical output of the liver, as measured by hippurate synthesis from benzoate, decreased from the control value of 98.4 to 32.7 mg/h. These results indicate that synergistic decreases in ADPf, Vmax, v and delta v/delta ADPf take place in the course of deterioration of mitochondrial ATP synthesis and work output in jaundiced liver.

[Nutritional and immunological assessment in patients with malignant obstructive jaundice--the influence of preoperative biliary decompression and abdominal surgery].

A clinical study was undertaken to evaluate the nutritional and immunological status of patients with malignant obstructive jaundice with serum bilirubin level of 5mg/dl or greater (N = 22). Healthy volunteers (N = 9) and patients with cholecystolithiasis (N = 26) were assessed as non-icteric control, as well as patients with obstructive jaundice due to choledocholithiasis (N = 3) were used as icteric control. We also evaluated the time-course of five patients with malignant obstructive jaundice before and after PTCD and surgery. Nutritional status was assessed by prognostic nutritional index (PNI), blood chemistry and rapid turnover protein. Cellular and humoral immunity were assessed by peripheral lymphocyte count, lymphocyte stimulation test, antibody dependent cell-mediated cytotoxicity, and serum immunoglobulins. The following results were obtained. 1) Nutritional and immunological status of patients with malignant obstructive jaundice were severely depressed. 2) Nutritional and immunological status were not improved by preoperative biliary decompression such as PTCD. 3) The immunological indices recovered postoperatively. However, the nutritional indices did not recover by the time of discharge. Intensive nutritional support before and after surgery may be mandatory for patients with malignant obstructive jaundice.

An assessment of percutaneous transhepatic biliary drainage in the correction of the metabolic capacity of the jaundiced liver by hippurate-synthesizing test.

Hippurate-synthesizing ability was investigated in patients with jaundice with percutaneous transhepatic biliary drainage (PTBD) in relation to hepatic metabolic capacity. In 16 patients with PTBD because of obstructive jaundice and 11 patients without hepatic disease, 1.77 grams of sodium benzoate was injected and the amount of hippurate synthesized and excreted in the urine collected at 30, 60, 120 and 180 minutes was measured (hippurate test). In patients with jaundice and patients in the control group, an almost linear increase was observed in the level of urinary hippurate after benzoate loading. However, the values of the patients with jaundice at one and two hours after the benzoate loading were significantly lower than those of the patients in the control group. Serum levels of glutamate oxaloacetic transaminase, glutamate pyruvate transaminase, alkaline phosphatase, gamma-glutamyl transpeptidase, total bilirubin and direct bilirubin were significantly decreased during PTBD (p < 0.05). Bilirubin levels were closely correlated with hippurate test values (r = 0.567, p < 0.05). Values were also correlated with the period of PTBD before the hippurate test was performed (r = 0.632, p < 0.05). Recovery in hippurate synthesizing ability was observed when the total bilirubin levels decreased to less than 5 milligrams per deciliter or PTBD was maintained for more than three weeks. Because hippurate synthesis is dependent on adenosine triphosphate supply in the hepatic mitochondria, the value of the hippurate test reflects the metabolic viability of the liver in relation to energy metabolism. It is also suggested that the steady maintenance of PTBD for three weeks or more with a decrease in total bilirubin level less than 5 milligrams per deciliter is necessary for full recovery of the metabolic capacity of the jaundiced liver.

Semiquantitative assessment of cholestasis and lymphocytic piecemeal necrosis in primary biliary cirrhosis: a histologic and immunohistochemical study.

Histologic activity of chronic cholestasis and lymphocytic piecemeal necrosis, a characteristic finding of chronic active hepatitis, was examined semiquantitatively in 157 liver biopsy specimens from 122 patients with primary biliary cirrhosis (PBC). Although both of these lesions were usually admixed variably in a single liver specimen, semiquantitative assessment made it possible to classify liver biopsy specimens into four groups: group A, no or minimum cholestatic or hepatitic changes (58 specimens); group B, predominantly cholestatic changes (37 specimens); and group C, predominantly hepatitic changes (54 specimens). Only eight specimens fell into group D, prominent cholestatic as well as hepatitic changes. Serial liver biopsies of specimens within groups B and C showed a persistence of group B- and C-type pathologies, while liver biopsies of group A specimens frequently changed to group B or C. Immunohistochemical studies illustrated that lymphocytic piecemeal necrosis mainly consisted of activated T lymphocytes as seen in chronic active hepatitis. Our data suggest that either of two hepatic parenchymal lesions predominates and persists in each liver biopsy specimen. A high cholestatic score appeared to relate to poor prognoses of the patients and also to the degree of cirrhotic transformation. This grouping system may be valuable in the clinicopathologic assessment of PBC, when it is combined with ordinary staging.

Sirius red histophotometry and spectrophotometry of sections in the assessment of the collagen content of liver tissue and its application in growing rat liver.

By means of staining with Sirius Red F3BA in a saturated picric acid solution, the collagen contents of rat livers with varying degrees of fibrosis have been measured quantitatively in fixed and sectioned material, using both histophotometry in situ and extraction of bound dye with colorimetric analysis. These findings have been correlated with chemical assays of the hydroxyproline content in homogenates from the same livers. It appears that a highly significant correlation exists between both section-based analysis methods and the hydroxyproline content, the Spearman-Rank correlation coefficients being virtually identical. For analysis of collagen accumulation in rat liver, both section-based methods seem to be useful and reliable, the extraction method giving the quickest results for large-scale screening, and the histophotometric method being more appropriate to take readings in selected areas. With human liver material, indications have been obtained for the existence of large sampling errors due to inhomogeneous distribution of collagen deposits. Using the extraction method, no significant changes could be observed in the volume density of collagen during postnatal growth from 1 week to 21 months in rat liver: only on the third day after birth was a higher value of collagen/total protein obtained, possibly due to a higher water content of the hepatocytes. Partial hepatectomy was found to have no influence at all on the collagen content of rat liver during the period of restorative growth or after it.

Quantitative assessment of hepatic function by breath analysis after oral administration of (14C)aminopyrine.

The rate of hepatic metabolism of dimethylaminoantipyrine (aminopyrine), which occurs primarily through N-demethylation, was assessed by measurement of the specific activity of 14CO2 excreted in breath samples obtained 2 hours after oral administration of a trace dose of [14C]aminopyrine. The percentage of administered 14C excreted in 14CO2 in 2 hours was 7.0 +/- 1.3 (SD)% in control patients, and significantly less (P less than 0.01) in patients with portal cirrhosis (2.6 +/- 1.2%), fatty liver (4.7 +/- 1.1%), hepatitis (2.6 +/- 1.4%), and hepatic malignancy (3.5 +/- 1.8%). In 16 of 24 subjects with cholestasis not caused by malignant disease the mean 14CO2 excretion was normal. The 14CO2 excretion in patients with portal cirrhosis correlated highly with aminopyrine metabolic clearance rate (r equals 0.92), serum albumin (r equals 0.75), and retention of bromsulphalein (r equals 0.73). Abnormal 14CO2 excretion returned to normal in patients with hepatitis, when the hepatitis resolved. The data suggest that the aminopyrine breath test is a safe, simple, qualitative and quantitative liver function test.