[Show abstract][Hide abstract] ABSTRACT: To investigate the significance of downregulation of liver fatty acid-binding protein (L-FABP) expression in hepatocellular carcinoma (HCC).
Tissue microarrays of 146 cases of HCC were used to perform immunohistochemical staining for L-FABP. For each L-FABP-negative HCC, further immunohistochemical staining was performed using a representative whole-tissue section to confirm the downregulation of L-FABP expression and to assess the intratumoral heterogeneity of the staining pattern. Clinical data were retrieved from the clinical files, and histological slides were reviewed. Immunohistochemical staining for cytokeratin (CK) 7, CK 19, β-catenin, glutamine synthetase (GS), and serum amyloid A were also performed on the tissue microarrays. Clinicopathological features of the L-FABP-negative and L-FABP-positive HCC cases were compared. Furthermore, L-FABP and GS gene expression in HCC and cholangiocarcinoma cell lines were analyzed using real-time reverse transcription polymerase chain reaction. Mutation analysis of HNF1A [encoding hepatocyte nuclear factor 1 (HNF1)α] was performed for L-FABP-negative HCC cases.
Sixteen (10.9%) of the 146 cases of HCC stained negative for L-FABP. When we examined the correlation between the downregulation pattern of L-FABP and tumor size, most cases of smaller HCC (≤ 2 cm in diameter) exhibited focal downregulation, while most cases of larger HCC (> 2 cm in diameter) exhibited diffuse downregulation. The correlation was statistically significant (P = 0.036). When the HCC was smaller, the L-FABP-negative area often corresponded to a "nodule-in-nodule" appearance. Among the small HCC cases, tumor differentiation was significantly lower, and the frequency of intratumoral inflammation was significantly lower in L-FABP-negative cases than in L-FABP-positive cases (P = 0.032 and P = 0.009, respectively). The frequency of positivity for β-catenin and GS staining was significantly higher in L-FABP-negative cases of small HCC than in L-FABP-positive cases of small HCC (P = 0.009 and P = 0.000, respectively). Among six HCC cell lines examined, four showed higher expression of L-FABP, and the remaining two cell lines showed lower or no expression of L-FABP. Two of the 16 L-FABP-negative HCC cases possessed a mutation in exon 4 of HNF1A.
In smaller HCC, L-FABP downregulation probably occurs because of phenotypic changes during tumor progression. Moreover, this downregulation correlated with tumor differentiation and intratumoral inflammation.
World journal of gastroenterology : WJG. 12/2014; 20(46):17541-51.
[Show abstract][Hide abstract] ABSTRACT: Recent studies have indicated that hepatocellular adenoma (HCA) is a heterogenous group of benign tumors with various genetic and clinicopathological characteristics.
We delineated the clinicopathological characteristics of HCA subtypes and evaluated the expression of transporter protein OATP1B3 in HCAs.
HCAs in 34 Japanese patients were investigated immunohistochemically and classified into four subtypes (HNF1α-inactivated type, H-HCA; β-catenin-activated type, b-HCA; inflammatory type, I-HCA; unclassified type, u-HCA). Immunostaining of OATP1B3 protein in HCA tissue sections was performed to determine the association between OATP1B3 expression and HCA subtypes.
HCA was categorized into the following four subtypes and two combined subtypes: 10 H-HCAs (29%), 10 I-HCAs (29%), 7 b-HCAs (21%), 2 b-HCA/H-HCA (6%), 2 b-HCA/I-HCA (6%), and 3 u-HCAs (9%). The male-to-female ratio was 18:16. Oral contraceptive use was rare but 7 HCAs were found in patients with glycogen storage disease, congenital absence of the portal vein, and idiopathic portal hypertension. OATP1B3 expression was decreased in 24 HCAs but was preserved or increased in 10 HCAs. All nine HCAs with nuclear staining for β-catenin showed preserved or enhanced OATP1B3 expression, indicating a significant association between nuclear β-catenin accumulation and OATP1B3 expression in HCAs.
HCA subtype classification was validated in 91% of our Japanese subjects although their clinical backgrounds including rare contraceptive use were different from European subjects. A close association between preserved or enhanced OATP1B3 expression and b-HCA subtype indicated important modalities for clinical decisions in the treatment and follow-up of patients with HCAs.
This article is protected by copyright. All rights reserved.
[Show abstract][Hide abstract] ABSTRACT: Nonalcoholic fatty liver disease (NAFLD), a hepatic manifestation of metabolic syndrome, is the most common chronic liver disease, and the prevalence is rapidly increasing worldwide. Nonalcoholic steatohepatitis (NASH), the severe form of NAFLD, can progress to liver cirrhosis and hepatocellular carcinoma (HCC). Although noninvasive clinical scores and image-based diagnosis for NAFLD have improved, histopathological evaluation of biopsy specimens remains the gold standard for diagnosing NAFLD/NASH. Steatosis, lobular inflammation, and hepatocellular ballooning are all necessary components for the diagnosis of NASH; fibrosis is also typically observed. Other histopathological abnormalities commonly observed in NASH include hepatocellular glycogenated nuclei, lipogranulomas, and acidophil bodies. The characteristics of pediatric NAFLD/NASH differ from adult NAFLD/NASH. Specifically, steatosis and portal inflammation are more severe in pediatric NAFLD, while intralobular inflammation and perisinusoidal fibrosis are milder. Although interobserver agreement for evaluating the extent of steatosis and fibrosis is high, agreement is low for intralobular and portal inflammation. A recently reported histological variant of HCC, steatohepatitic HCC (SH-HCC), shows features that resemble non-neoplastic steatohepatitis, and is thought to be strongly associated with underlying NASH. In this report, we review the histopathological features of NAFLD/NASH.
World Journal of Gastroenterology 11/2014; 20(42):15539-15548. · 2.43 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: AimsWe characterized serum amyloid A (SAA)-positive hepatocellular neoplasm/nodules arising in alcoholic cirrhosis, which is detected as a hypervascular hepatocellular nodules resembling hepatocellular carcinoma on imaging.Methods and ResultsFifty-three hepatocellular nodules were examined using immunostaining for SAA, glutamine synthetase and glypican-3 in 23 patients (4 women and 19 men) with alcoholic cirrhosis. Sixteen nodules were examined on the magnetic resonance imaging with EOB enhancement (EOB-MRI). Somatic mutations on IL6ST, GNAS and STAT3 were examined in 19 nodules. Thirty-six nodules in 18 patients were diagnosed as SAA-positive hepatocellular neoplasm/nodules and the remaining seventeen nodules in 8 patients were SAA-negative focal nodular hyperplasia (FNH)-like nodules. SAA-positive hepatocellular neoplasms/nodules showed significantly more extensive sinusoidal dilatation, inflammatory reaction, abnormal thick arteries and cellular atypia than FNH-like nodules (p<0.05). Eight SAA-positive hepatocellular neoplasms/nodules (67%) showed slight hypointensity in the hepatobiliary phase on the EOB-MRI, whereas all 4 FNH-like nodules showed iso-intensity (p<0.05). STAT3 mutations were detected in 2 of 17 SAA-positive hepatocellular neoplasms/nodules.Conclusions
This study disclosed that about two thirds of hypervascular hepatocellular nodules arising in alcoholic cirrhosis were SAA-positive hepatocellular neoplasms/nodules which show different finings on the EOB-MRI. STAT3 mutations were detected in 11.8% of SAA-positive hepatocellular neoplasms/nodules, supporting neoplastic nature.This article is protected by copyright. All rights reserved.
[Show abstract][Hide abstract] ABSTRACT: Few studies have investigated the effects of Japanese herbal medicines on nonalcoholic fatty liver disease (NAFLD)/nonalcoholic steatohepatitis (NASH). To the best of our knowledge, only one study has examined whether high-fat (HF) diet-fed db/db mice are appropriate animal models of NASH. We investigated the effects of four types of Japanese herbal medicines (shosaikoto (TJ-9), inchinkoto (TJ-135), juzentaihoto (TJ-48), and keishibukuryogan (TJ-25)) on hepatic lesions of HF diet-fed db/db mice. Db/db mice were divided into six groups: control diet (control); HF diet (HF); and HF diet supplemented with TJ-9, TJ-135, TJ-48, or TJ-25 (TJ-9, TJ-135, TJ-48, and TJ-25, respectively). Mice were killed after 6 weeks of treatment, and biochemical and pathological analyses were performed. Mice in the HF group consistently developed histopathological features consistent with definite NASH, and marked necroinflammation occurred. Serum alanine aminotransferase levels in the TJ-9, TJ-135, and TJ-48 groups were significantly improved compared with those in the HF group. With regard to liver histology, TJ-9 and TJ-48 significantly improved lobular inflammation, and TJ-135 significantly improved ballooning degeneration. We have shown that HF diet-fed db/db mice are animal models that correctly recapitulate the histopathology of human NASH and that TJ-9, TJ-135, and TJ-48 inhibit necroinflammatory activity in this model.
Pathology International 10/2014; · 1.59 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background
Gastric cancer (GC) is a common cancer globally. miRNA-21 (miR-21) appears to be important in the tumourigenesis of almost all types of human cancer. However its precise localization in GC has not yet to be clarified. We thus examined miR-21 localization in GC and revealed its clinicopathological importance.Methods
Tissue arrays of 469 GCs from 454 patients were examined for miR-21 using in situ hybridization (ISH). The positivity was evaluated separately in tumour cells and stromal cells. Conventional sections of 10 GCs were also stained. Eight cases were examined by quantitative RT-PCR (qRT-PCR).ResultsmiR-21 was highly expressed in tumour cells of 44% of cases and in cancer stroma of 51% of cases. miR-21 of tumour cells was not related to clinicopathological factors, whereas stromal miR-21 was related to many factors including tumour stage, size, and nodal metastasis. Stromal miR-21 gradually increased during tumour progression. ISH of whole sections showed stronger stromal positivity in invasive areas with desmoplastic reaction. Cancer stroma also showed higher miR-21 expression than tumour and non-tumourous tissue in the qRT-PCR study.Conclusion
Stromal miR-21 is closely related to tumour progression in GC. Stromal miR-21 of tumours might be a target of treatment.This article is protected by copyright. All rights reserved.
[Show abstract][Hide abstract] ABSTRACT: The aim of this study was to evaluate integrin expression and immunolocalization of the extracellular matrix proteins in cholangiolocellular carcinoma (CoCC).
Tissue specimens of 23 CoCCs, 28 cholangiocarcinomas (CCCs), 42 hepatocellular carcinomas (HCCs), and 11 classical type combined hepatocellular-cholangiocarcinomas (CHCs) were immunostained for β6, β4 and α3 integrins, fibronectin and laminin. ITGB6, B4 and A3 mRNA levels in six HCC cell lines, five CCC cell lines and two CHC cell lines were quantified by quantitative reverse transcription-polymerase chain reaction.
Little or no positivity for β6, β4 and α3 integrins was shown in 91%, 91% and 52% of CoCCs and 100%, 98% and 81% of HCCs, respectively, according to immunostaining, whereas intense positive staining for these integrins was demonstrated in 64%, 96% and 75% of CCCs, respectively. There was a close correlation between β4 and α3 integrin expression and intracytoplasmic laminin in CoCC, CCC and HCC, but not between β6 expression and its ligand, fibronectin. Integrin mRNA levels were increased in four of five CCC cell lines, but nearly undetectable in five of six HCC cell lines and one CHC cell line. Tubular differentiation of a CHC cell line cultured in collagen-gel matrix induced up-regulation of these integrins.
Our results first indicated down-regulation of αvβ6, α6β4 and α3β1 integrins in CoCC, in contrast to its high expression in CCC, suggesting a diagnostic value of integrins in the differential diagnosis of CoCC and CCC, as well as a useful inducible marker of the intermediate features of CoCC.
[Show abstract][Hide abstract] ABSTRACT: Although Japanese herbal medicines (JHMs) are widely used in Japan, only a few studies have investigated their effects on nonalcoholic steatohepatitis (NASH). In the present study, we examined the effect of 4 kinds of JHMs [sho-saiko-to (TJ-9), inchin-ko-to (TJ-135), juzen-taiho-to (TJ-48), and keishi-bukuryo-gan (TJ-25)] on a mouse model of NASH. Db/db mice were divided into 6 groups: control diet (control), methionine- and choline-deficient diet (MCD), and MCD diet supplemented with TJ-9, TJ-135, TJ-48, and TJ-25 (TJ-9, TJ-135, TJ-48, and TJ-25, respectively). All mice were sacrificed after 4 weeks of treatment, and biochemical, pathological, and molecular analyses were performed. Serum alanine aminotransferase levels and liver histology, including necroinflammation and fibrosis, were significantly alleviated in the TJ-9 and TJ-48 groups compared with the MCD group. The expression level of transforming growth factor (TGF)-β1 mRNA in the liver was significantly suppressed by TJ-48. Although the differences were not statistically significant, the expression levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 were lower, and those of peroxisome proliferators-activated receptor (PPAR)γ were higher in the TJ-9 and/or TJ-48 groups than in the MCD group. Similarly, even though the results were not statistically significant, malondialdehyde levels in liver tissues were lower in the TJ-9 and TJ-48 groups than in the MCD group. We showed that JHMs, especially TJ-9 and TJ-48, inhibited the necroinflammation and fibrosis in the liver of a mouse model of NASH, even though the mechanisms were not fully elucidated. Further studies are needed in the future to investigate the possibility of clinical application of these medicines in the treatment for NASH.
PLoS ONE 01/2014; 9(1):e87279. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Flat epithelial lesions of the biliary tract cannot be detected by the image analysis, and the diagnosis entirely depends on pathological examination. The biliary tract is often affected by inflammatory conditions, and the resultant changes of the biliary epithelium make it difficult to differentiate them from neoplasia. Thus, the pathological diagnosis of biliary flat epithelial lesions can be challenging. In the biliary tract, there are several forms of intraepithelial neoplasia of the flat type, and biliary intraepithelial neoplasia (BilIN) is known as one of such lesions that represent the multistep cholangiocarcinogenesis. In this article, the diagnostic criteria and the differential diagnosis of biliary flat epithelial lesions, particularly focusing on BilIN, were presented and discussed to provide help to advance clinical and research applications of the BilIN system.
Journal of Gastroenterology 01/2014; · 4.02 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Great progress has been made in the diagnosis of focal nodular hyperplasia (FNH) and hepatocellular adenoma (HCA) in the last few years due to the use of molecular criteria. This has allowed us to identify a new type of hepatic nodule. In this case report, we present a male patient with a hepatic nodule associated with idiopathic portal hypertension (IPH) pathologically exhibiting not only the morphological features of FNH, such as ductular reactions, dilated sinusoids, major vascular abnormalities and an immunohistochemical "map-like" pattern of glutamine synthetase (GS), but also the immunohistological features of focal HCA, such as strong expression of serum amyloid A and C-reactive protein and weak expression of GS. As the final diagnosis, the nodule was identified as an FNH-like lesion with focal inflammatory hepatocellular adenoma.
[Show abstract][Hide abstract] ABSTRACT: The estrogen receptor (ER) is a key molecule for growth of breast cancers. It has been a successful target for treatment of breast cancers. Elucidation of ER expression mechanism is of importance for designing therapeutics for ER-positive breast cancers. However, the detailed mechanism of ER stability is still unclear. Here we report that histone acetyltransferase Hbo1 promotes destabilization of estrogen receptor α (ERα) in breast cancers through lysine 48-linked ubiquitination. The acetyltransferase activity of Hbo1 is linked to its activity for ERα ubiquitination. Depletion of Hbo1 and anti-estrogen treatment displayed a potent growth suppression of breast cancer cell line. Hbo1 modulated transcription by ERα. Mutually exclusive expression of Hbo1 and ERα was observed in roughly half of human breast tumors. Modulation of ER stability by Hbo1 in breast cancers may provide a novel therapeutic possibility. This article is protected by copyright. All rights reserved.
[Show abstract][Hide abstract] ABSTRACT: Epigallocatechin 3-gallate (EGCG) has cytotoxic effects in many cancer cells. It has been reported that A549 lung cancer cells are markedly resistant to cell death induced by EGCG. In the present study, the effects of EGCG on A549 lung cancer cell growth and angiogenesis were studied. We found that EGCG dose-dependently suppressed A549 cell growth, while A549 cells were markedly resistant to cell death in vitro. Next we next found that EGCG increased endostatin expression and suppressed vascular endothelial growth factor (VEGF) expression. We further studied to determine whether EGCG would suppress A549 tumor growth in nude mouse and angiogenesis. EGCG in drinking water significantly suppressed A549 tumor growth in nude mice. Histological analysis revealed that the number of CD34 positive vessels had a tendency to decrease in the tumor. In sum, EGCG had anti-proliferative effects of A549 on tumor growth and showed a tendency to suppress angiogenesis.
Bioscience Biotechnology and Biochemistry 09/2013; · 1.27 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In order to evaluate and judge a fibrotic stage of patients with chronic hepatitis C, multivariate regression analysis was performed using multiple fibrosis markers.
A total of 581 patients from 8 Hepatology units and institutes were diagnosed by needle biopsy as having chronic liver disease caused by hepatitis C virus. Twenty-three variables and their natural logarithmic transformation were employed in the multivariate analysis.
Multivariate regression analysis finally obtained the following function: z=2.89 x ln(type IV collagen 7S)(ng/ml) - 0.011 x (platelet count)(x10(3) /mm(3) ) + 0.79 x ln(total bilirubin)(mg/dl) + 0.39 x ln(hyaluronic acid)(microgram/L) - 1.87. Median values of the "fibrosis score" of F1 (N=172), F2 (N=80), F3 (N=37), and F4 (N=16) were calculated as 1.00, 1.45, 2.82, and 3.83, respectively. Multiple regression coefficient and coefficient of determination were 0.56 and 0.320, respectively. Validation with patient data from other institutions demonstrated good reproducibility of the "fibrosis score" for hepatitis C (FSC), showing 1.10 in F1 (N=156), 2.35 in F2 (N=73), 3.16 in F3 (N=36), and 3.58 in F4 (N=11), respectively.
A concise multiple regression function using four laboratory parameters successfully predicted pathological fibrosis stage of patients with hepatitis C virus infection. (195 words).
[Show abstract][Hide abstract] ABSTRACT: Twelve hepatocellular nodules were characterized in the resected livers from 3 patients (2 men and a woman) with alcoholic cirrhosis. Imaging techniques suggested that the nodules were hypervascular and may be hepatocellular carcinoma. Five nodules (4-31mm in diameter) were serum amyloid A-positive hepatocellular neoplasm, which shares features with inflammatory hepatocellular adenoma. The remaining 7 nodules (5-8mm) were focal nodular hyperplasia-like nodules showing focal or no immunostaining for serum amyloid A. The serum amyloid A-positive hepatocellular neoplasms showed increased cellular density, inflammatory infiltrate, sinusoidal dilatation, and ductular reaction to various degrees. These histologic features tended to be less extensive in focal nodular hyperplasia-like nodules. Three of 4 serum amyloid A-positive hepatocellular neoplasms showed slight hypointensity in the hepatobiliary phase on the magnetic resonance (MR) imaging with gadolinium ethoxybenzyl diethylenetriaminepentaacetic acid (Gd-EOB-DTPA) enhancement. In contrast, 3 focal nodular hyperplasia-like nodules showed iso-intensity in the hepatobiliary phase. This study further confirms characteristics of serum amyloid A-positive hepatocellular neoplasm arising in alcoholic cirrhosis that share features with inflammatory hepatocellular adenomas. Serum amyloid A-positive hepatocellular neoplasms sometimes co-exist with focal nodular hyperplasia-like nodules and may show different findings on Gd-EOB-enhanced MR imaging.
Histology and histopathology 05/2013; · 2.24 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: AIM: The aim of this study was to investigate the expression of farnesoid X receptor (FXR) in human hepatocellular carcinoma (HCC) tissues and cell lines and evaluate its clinicopathological significance. METHODS: Expression levels of FXR protein and mRNA in hepatocytes, hepatic stellate cells (SC), and HCC cells in human liver, HCC tissues and cultured cell lines were analyzed using immunohistochemical methods, western blotting (WB), and quantitative reverse transcription polymerase chain reaction (qRT-PCR). The relationship between FXR expression and clinicopathological parameters was also investigated. RESULTS: Immunoreactivity for FXR was observed in nuclei of hepatocytes, SC and HCC cells. The intensity of nuclear FXR positive staining was comparable or increased in tumor cells of all HCC tissues when compared with hepatocytes of non-tumorous liver tissues of the same patients as well as in comparison with metastatic colon cancers. A significant level of FXR expression in four of six human HCC cell lines was also confirmed, while it was undetectable in three cholangiocarcinoma cell lines. However, FXR protein and mRNA levels in HCC tissues determined by WB and qRT-PCR were lower than those in non-tumorous liver tissues because of the high level of FXR expression in SC nuclei as detected by immunohistochemical double stain. Statistically significant relationships between FXR immunostaining intensity and high Ki-67 labeling indices or a history of transcatheter arterial chemoembolization in HCC patients were also disclosed. CONCLUSION: Contrary to previous reports, preserved or enhanced expression levels of nuclear FXR were detected in HCC, indicating that FXR may play significant roles in the biological behavior of HCC.
[Show abstract][Hide abstract] ABSTRACT: Russell bodies are globular and eosinophilic inclusion bodies in the cytoplasm of mature plasma cells. Plasma cells whose cytoplasm is filled with Russell bodies are designated as Mott cells. Russell body duodenitis (RBD) is a unique form of chronic duodenitis that is characterized by infiltration of numerous Mott cells. RBD is very rare; only two cases have been reported to date. In this paper, we report a case of RBD in a patient with retroperitoneal metastasis of ureteral cancer. A 77-year-old man was admitted to our hospital complaining of appetite loss, vomiting, and upper abdominal distension. He had undergone left nephroureterectomy for ureteral cancer 4 years earlier. Upper digestive tract endoscopy revealed edema, stenosis, and punctate redness of the mucosa of the duodenum, and a biopsy was performed. Histological analysis showed that numerous Mott cells had infiltrated the lamina propria mucosae, and the condition was diagnosed as RBD. A mass lesion in the retroperitoneum adjacent to the duodenum was detected by abdominal computed tomography, and was diagnosed as metastatic urothelial carcinoma by biopsy. It is possible that chemokines produced by tumor cells caused RBD in this case.
World Journal of Gastroenterology 01/2013; 19(1):125-8. · 2.43 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: AimThe aim of this study was to investigate the expression of farnesoid X receptor (FXR) in human hepatocellular carcinoma (HCC) tissues and cell lines and evaluate its clinicopathological significance. Methods
Expression levels of FXR protein and mRNA in hepatocytes, hepatic stellate cells (SC), and HCC cells in human liver, HCC tissues and cultured cell lines were analyzed using immunohistochemical methods, western blotting (WB), and quantitative reverse transcription polymerase chain reaction (qRT–PCR). The relationship between FXR expression and clinicopathological parameters was also investigated. ResultsImmunoreactivity for FXR was observed in nuclei of hepatocytes, SC and HCC cells. The intensity of nuclear FXR positive staining was comparable or increased in tumor cells of all HCC tissues when compared with hepatocytes of non‐tumorous liver tissues of the same patients as well as in comparison with metastatic colon cancers. A significant level of FXR expression in four of six human HCC cell lines was also confirmed, while it was undetectable in three cholangiocarcinoma cell lines. However, FXR protein and mRNA levels in HCC tissues determined by WB and qRT–PCR were lower than those in non‐tumorous liver tissues because of the high level of FXR expression in SC nuclei as detected by immunohistochemical double stain. Statistically significant relationships between FXR immunostaining intensity and high Ki‐67 labeling indices or a history of transcatheter arterial chemoembolization in HCC patients were also disclosed. Conclusion
Contrary to previous reports, preserved or enhanced expression levels of nuclear FXR were detected in HCC, indicating that FXR may play significant roles in the biological behavior of HCC.
Hepatology Research 01/2013; 43(9). · 2.22 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: AIM: In order to evaluate and judge a fibrotic stage of patients with chronic hepatitis B, multivariate regression analysis was performed using multiple fibrosis markers. METHOD: A total of 227 patients from seven hepatology units and institutes were diagnosed by needle biopsy as having chronic liver disease caused by hepatitis B virus. Twenty-three variables and their natural logarithmic transformation were employed in the multivariate analysis. Multiple regression function was generated from data of 158 patients in one hospital, and validation was performed using the other data of 69 patients from six other hospitals. RESULTS: After stepwise variable selection, multivariate regression analysis finally obtained the following function: z = 1.40 × ln (type IV collagen 7S) (ng/mL) - 0.017 × (platelet count) (×1000(3) /mm(3) ) + 1.24 × ln (tissue inhibitor of matrix metalloproteinase-2) (ng/mL) + 1.19 × ln (α-2-macroglobulin) (mg/dL) - 9.15. Median values of fibrosis scores of F1 (n = 73), F2 (n = 42), F3 (n = 31) and F4 stages (n = 12) were calculated as 0.95, 2.07, 2.98 and 3.63, respectively. Multiple regression coefficient and coefficient of determination were 0.646 and 0.418, respectively. Validation with patient data from other institutions demonstrated good reproducibility of fibrosis score for hepatitis B (FSB), showing 1.33 in F1 (n = 27), 2.20 in F2 (n = 20), 3.11 in F3 (n = 20) and 5.30 in F4 (n = 2), respectively. CONCLUSION: A concise multiple regression function using four laboratory parameters successfully predicted pathological fibrosis stage of patients with hepatitis B virus infection.