HCC-ART score, a simple, highly sensitive and specific test for early diagnosis of hepatocellular carcinoma: a large-scale, multicentre study.

BACKGROUND: A simple scoring system is needed to discriminate HCC from patients with chronic liver diseases (CLD). The simplest score would be one that requires only variables that can be documented simply from routine laboratory tests without the need for sophisticated tests. METHODS: Data from the estimation group (1351 patients) and the validation group (2208 patients) were retrospectively analysed. Liver fibrosis-negative control and liver cirrhosis were compared with HCC. Area under ROC curve (AUC) were used to develop HCC-α-fetoprotein-routine test (HCC-ART). RESULTS: Hepatocellular carcinoma-AFP-routine test showed diagnostic accuracy for liver cirrhosis vs HCC with ROC curves of 0.99%, sensitivity of 97%, and specificity of 96% in the estimation, and 0.95%, 90%, and 83%, respectively, in the validation. Sensitivity (97%) and specificity (100%) were obtained to discriminate HCC from liver fibrosis. Area under curve for AFP at 400 U l(-1) was 0.70, sensitivity was 41%, and specificity was 99% in the estimation, and 0.77%, 54%, and 99%, respectively, in the validation. The AUC for HCC-ART in HCC with single tumour, absent vascular invasion, size <2 cm and CLIP score (0-1) were 0.95, 0.93, 0.86, 0.87, respectively, compared with 0.72, 0.71, 0.71, 0.50, respectively, for AFP. CONCLUSION: Hepatocellular carcinoma-AFP-routine test could increase the accuracy of HCC screening and surveillances and could be used worldwide without extra efforts.

Evaluation of serum procollagen aminoterminal propeptide III, laminin, and hydroxyproline as predictors of severe fibrosis in patients with chronic hepatitis C.

In an attempt to identify biochemical analytes that could enhance the discrimination between the patients with severe liver fibrosis (F3-F4) and mild fibrosis (F1-F2) based on absolute values of biochemical markers, we measured 12 analytes, including procollagen III aminoterminal propeptide (PIIINP), laminin, proline, hydroxylproline, glycine, AST, ALT, alkaline phosphatase, albumin, total bilirubin, total protein, and prothrombin time in 252 individuals with chronic hepatitis C infection (CHC). PIIINP and laminin were determined by radio-immunoassay; the degraded amino acids were determined using high performance liquid chromatography. Statistical analyses were performed by logistic regression, and receiver operating characteristic (ROC) curves. The best linear combination of blood markers was selected by multivariate discriminant analysis (MDA) for construction of the fibrosis discriminant score (FDS). FDS, an index of five markers (PIIINP, laminin, hydroxyproline, prothrombin activity, and AST/ALT) correctly classified 82% of the patients with severe liver fibrosis at a discriminant cut-off score=-0.5 (i.e., less than -0.5 indicated severe liver fibrosis and greater than -0.5 indicated mild liver fibrosis with sensitivity (76%) and specificity (89%). This result was reproduced in a validation study with no significant difference. In conclusion, FDS is useful for identifying severe liver fibrosis in patients with CHC.

Dysregulation of blood lymphocyte subsets and natural killer cells in schistosomal liver cirrhosis and hepatocellular carcinoma.

Immunological factors are important in the pathogenesis of a wide spectrum of hepatobiliary diseases. Using flow cytometry, we determined the changes in lymphocyte subsets and natural killer cells in 123 individuals (81 patients with liver disease and 42 healthy volunteers). The liver diseases included periportal fibrosis (PPF, 10 patients), liver cirrhosis (LC, 31 patients), and hepatocellular carcinoma (HCC, 40 patients). Schistosomiasis and viral hepatitis B and C were the putative etiological agents of liver diseases. Immunophenotyping by indirect immunofluorescence was conducted using monoclonal antibodies to CD3 (T-lymphocytes), CD4 (helper/inducer T-cells), CD8 (suppressor/cytotoxic T-cells), and CD57 (natural killer cells) cell surface markers. Immunophenotyping of PPF patients showed no significant changes in all markers compared with the healthy controls. However, there was a significant decrease ( P<0.01) in CD3 and CD4 T-cells, and a highly significant increase ( P<0.001) in CD57 T-cells in patients with LC or HCC. In addition, LC and HCC patients showed no significant change in CD8 T-cells compared with controls. In conclusion, the progression of liver diseases is associated with a dysregulation of cellular immune responses. T-lymphocytes and natural killer cells may play a role in the immunopathogenesis of liver cirrhosis and HCC.

Identification and characterization of a 26- to 28-kDa circulating antigen of Fasciola gigantica.

As a disease of domestic ruminants, fascioliasis is of considerable economic importance. Although serological tests are available for the diagnosis of the disease, they are of generally low specificity because of cross-reactivity with antigens from other parasites. There is a need to identify other Fasciola antigens on which more specific tests could be based. In the present study, a specific rabbit anti-serum and western-blot analyses were used to demonstrate the presence of a highly reactive antigen of 26-28 kDa not only in an extract of adult F. gigantica but also in the excretory/secretory products of the worms and in the bile secretions and sera of cattle that were naturally infected with this parasite. The 26- to 28-kDa antigen was isolated from preparative polyacrylamide gels, by electro-elution. The purified antigen showed a single peak at 5.8 min when analysed by capillary zone electrophoresis. It was characterized as protein containing 47.5% hydrophilic and 29.3% hydrophobic amino acids. Immunostaining demonstrated that the target epitope was located in the gut and tegument of adult F. gigantica and within the bile ducts, the portal tracts of the livers and the mucosa and muscularis of the gallbladders of infected cattle. A simple and rapid dot-ELISA technique based on the specific rabbit anti-serum was 100% specific when tested on the sera from nine cattle infected with F. gigantea and 27 uninfected cattle. In conclusion, the 26- to 28-kDa Fasciola antigen may be a promising candidate for the immunodiagnosis of fascioliasis.

DNA ploidy of liver biopsies from patients with liver cirrhosis and hepatocellular carcinoma: a flow cytometric analysis.

Flow cytometric DNA analysis was used to assess cellular kinetics of needle liver biopsies from patients with liver cirrhosis and hepatocellular carcinoma (HCC). An abnormal DNA content was shown in 44.5% of liver cirrhosis cases and in 78.6% of tumor sites. The number of proliferating cells (S + G2M%) was significantly increased in cirrhotic liver (P < 0.05). Dysplasia was found in 66% of cirrhotic specimens. All negative dysplasia specimens showed a diploid pattern while 69% of positive dysplastic specimens were aneuploid (P < 0.001). In conclusion, cell proliferation, aneuploidy and liver cell dysplasia are important indicators in liver cirrhosis for the development of HCC.

Orthotopic bladder substitutes: histopathologic risk factors.

Prostate glands from 150 patients with carcinoma of the bilharzial bladder who underwent cystoprostatectomy were studied histopathologically by step sections. Prostatic urethral involvement by urothelial carcinoma was noted in 13 out of 96 (13.5%) and 5 out of 40 (12.5%) squamous and transitional cell tumors, respectively. None of the 12 adenocarcinomas and the two undifferentiated tumors showed involvement. Prostatic urethral involvement was as high as 19% in basal tumors and 26.7% in multifocal tumors compared to only 6.5% when the tumors occupied the bladder body. There was a significant increase in the incidence of prostatic urethral involvement from 9.5 to 35% when the prostate gland was involved. Prostate gland was involved in 20 out of 150 (13.3%). The bladder tumor was basal and infiltrating the prostate in 18 such cases. Seminal vesicles were infiltrated in 6 cases from the adjacent basal bladder tumors. We conclude that patients with basal or multifocal tumors are risky regarding bladder substitution and we recommend routine diagnostic transurethral prostatic biopsies and frozen sections from the site of urethral transection during cystoprostatectomy whenever bladder substitution controlled by the urethral sphincter is considered.

Elevation of glutathione, glutathione-reductase and nucleic acids in both normal tissues and tumour of breast cancer patients.

The present study was carried out to determine the variation in reduced glutathione (GSH), glutathione-reductase (GSH-R), DNA and RNA contents between tumours and adjacent normal tissue surgically removed from patients with breast cancer. A highly significant increase in the content of these parameters was found in tumours compared with adjacent normal breast tissue. Such observations were also found in the whole-blood GSH, GSH-R and leukocyte DNA and RNA content of these patients compared with that of normal women. The present data may provide relevant information about the behaviour of DNA, RNA, GSH and GSH-R in breast tumours that may be valuable in the chemotherapeutic modalities and prognosis of such tumours.

The immunopathology of Schistosoma mansoni granulomas in human colonic schistosomiasis.

The immunopathology of Schistosoma mansoni infection was studied in colonic biopsies obtained from 14 patients with established schistosomiasis. The characteristic lesions of this parasitic infection are mainly induced by the presence of living eggs in the tissue. Different types of lesions can be present simultaneously. The earliest lesions contain T-lymphocytes as well as accessory cells around living eggs. They transform into granulomas composed of eosinophils, T-lymphocytes, a few B-lymphocytes and large mononuclear cells expressing major histocompatibility (MHC) class II antigens. These cells are also Mac 387 positive. This means that they are monocytes/macrophages freshly recruited from the blood. In other, probably older, granulomas, MHC class II positive cells tend to disappear and the centrally located multinucleated giant cells are negative for antibodies directed against MHC class II antigens. It appears thus that the composition of the granulomas in schistosomiasis is variable. The lesions may have characteristics of cell-mediated immunity and/or of a foreign-body reaction. Contrary to what is often seen in Crohn's disease or intestinal tuberculosis no major hyperplasia of the lymphoid tissue is observed in the colon in association with S. mansoni infection.

Hydrolytic enzymes in normal and neoplastic human bladder tissue.

The value of hydrolytic enzymes in 29 specimens of human bladder cancer has been assessed. Acid and alkaline phosphatases, gamma-glutamyl transferase, arylsulphatases and lactate dehydrogenase were estimated in human bladder tumours and adjacent non-malignant tissue. Elevated enzyme levels were observed in the tumours rather than in the non-malignant tissue, except for adenocarcinoma. These increased levels were also noted in bilharzial bladder carcinoma but not in lymph node metastasis. Increased enzyme activity could be due to over-production by the tumour cells. It was concluded that the increase was associated primarily with the presence of tumours and the assay of these enzymes in biopsy specimens may have significant diagnostic and therapeutic implications.

Immunofluorescent localization of Schistosoma mansoni circulating cathodic antigen in tissues of infected mice using monoclonal antibody.

In the present study the kinetics of the uptake and deposition of the major circulating cathodic antigen (CCA) of Schistosoma mansoni in liver, spleen, and kidney of S. mansoni infected Swiss mice was investigated in relation to the duration of infection and infection dose (50, 100, 200 cercariae). The presence of antigen was studied with a direct immunofluorescence reaction on frozen sections of the mouse organs, using a fluorescein isothiocyanate (FITC)-labelled mouse IgM monoclonal antibody recognizing a repeating epitope of CCA. CCA was demonstrable from 2 weeks post infection (p.i.) onwards in Kupffer cells in the liver, from 3-4 weeks p.i. onwards in macrophages in the marginal zones in the spleen and from 8 weeks p.i. onwards in kidney glomeruli. The immunofluorescence reactions on CCA in kidney glomeruli, however, remained relatively weak.

Presence of Schistosoma mansoni antigens in liver, spleen and kidney of infected mice: a sequential study.

In the present study the kinetics of the uptake and deposition of Schistosoma mansoni antigens in liver, spleen and kidney of S. mansoni infected Swiss mice have been investigated in relation to duration of infection and infection dose (50, 100, 200 cercariae). The presence of antigen was studied with a direct immunofluorescence reaction on frozen sections of the organs, using a number of fluorescein isothiocyanate (FITC)-labeled antisera produced against various antigen preparations isolated from different life-cycle stages of the parasite. The presence of antigen was demonstrable with two of the antisera, directed against the circulating anodic antigen (CAA) and against total soluble egg antigen (SEA). CAA was demonstrable from 1 week post infection (p.i.) onwards in Kupffer cells in the liver, from 2-3 weeks p.i. onwards in macrophages in the marginal zones in the spleen and from 3 weeks onwards in kidney glomeruli. Immunofluorescence reactions on CAA in kidney glomeruli, however, were only weak positive until 12 weeks p.i., whereafter strong positive reactions were found. SEA was demonstrable from 5 weeks p.i. onwards in Kupffer cells in the liver and from 4 weeks p.i. onwards in macrophages of the spleen. In contrast to CAA, SEA was not detectable in kidney glomeruli.