Normal Hepatic Tissue Research Articles

Saline conducted electric coagulation (SCEC): original experience in experimental hepatectomy

To evaluate the feasibility and superiority of a new coagulating and hemostatic method named "saline conducted electric coagulation (SCEC)". The Peng's multifunction operative dissector (PMOD) was modified to enable saline to effuse persistently out of its nib at a constant speed. In a group of six New Zealand rabbits, two hepatic lobes of each rabbits were resected respectively by SCEC and conventional electric coagulation (EC). The features of SCEC were recorded by photo and compared with conventional EC. After 7 d, the coagulating depth was measured in each residual hepatic lobe. Hepatic tissue was dyed by hematoxylin and eosin (HE) and studied under a microscope. The coagulating depth increased with the continuation of SCEC time. Hepatectomies were performed successfully, no rabbit died in the perioperative period. The incisal surface of SCEC was gray-white with no red bleeding point. There was a thick solidified layer at the margin and a thin red-white intermittent layer between the solidified layer and normal hepatic tissue at the vertical section of SCEC. The mean coagulating depth of SCEC was 1.8 cm vs. 0.3 cm of conventional EC. Pathological examination showed a mild inflammatory reaction by SCEC. SCEC is a feasible and safe method for surgical hemostasis. As a new technique for liver resection, SCEC shows better coagulating effect and milder inflammatory reaction than conventional EC. Our study shows bloodless liver resection can also be performed by SCEC, especially for liver malignant tumor.

Positron Emission Tomography of Human Hepatocellular Carcinoma Xenografts in Mice Using Copper (II)-64 Chloride as a Tracer with Copper (II)-64 Chloride

The aim of this study was to assess copper metabolism of human hepatocellular carcinoma (HCC) with positron emission tomographic (PET) imaging using copper (II)-64 chloride ((64)CuCl(2)) as a tracer. PET imaging of athymic mice (n = 5) bearing extrahepatic HCC xenografts was performed 24 hours after the intravenous injection of (64)CuCl(2), followed by exvivo tissue radioactivity assay. Expression of human copper transporter 1 (hCTR1) in HCC cells and tissues was examined by real-time reverse transcription polymerase chain reaction and immunohistochemistry analysis, respectively. The extrahepatic HCC xenografts in mice with increased uptake of (64)Cu radionuclide were visualized on the micro-PET images obtained 24 hours after the intravenous injection of (64)CuCl(2). PET quantitative analysis revealed increased (64)Cu radioactivity in tumor tissues (2.7 ± 0.6 %ID/g) compared to that in the soft tissue of the left shoulder opposite to the tumor site (0.6 ± 0.2 %ID/g) and the brain (0.7 ± 0.1 %ID/g) but lower than that of the liver (16.6 ± 1.3 %ID/g). Expression of hCTR1 in the HCC cells and xenograft tumor tissues was demonstrated by real-time reverse transcription polymerase chain reaction and immunohistochemistry analysis, respectively. The expression level of hCTR1 in the Hep3B HCC xenograft tissues was lower than that detected in the normal hepatic tissues and the tissue samples of well-differentiated primary HCC. Variable expression of hCTR1 was detected in the tissue samples of moderately differentiated primary HCC. Extrahepatic human HCC xenografts in mice could be localized with (64)CuCl(2) PET imaging, which might be useful for the localization and quantitative assessment of copper metabolism in extrahepatic metastases of HCC in humans.

The expression and significance of signal regulatory protein a1 in autoimmune hepatitis

To observe the expression change of signal regulatory protein alpha1 (SIRPalpha1) in autoimmune hepatitis (AIH) and approach the relationship between SIRPalpha1 and the extent of inflammation. Immunohistochemistry is used to detect the expression of SIRPalpha1 in the paraffin section preparations of 33 AIH and 10 normal hepatic tissue. SIRPalpha1 is positive or weakly positive expressed in AIH. The staining is localized in the cytoplasm of Kupffer cells in the hepatic sinusoid with focal distribution. It is negative in normal hepatic tissue. In light AIH, it is negative or weakly positive expressed with a 36.4 percent of the positive rate (4/11). The positive or strong positive expression is found in the moderate AIH with an 84.2 percent of the positive rate(16/19). There is statistical significance between both light AIH, moderate AIH and severe AIH (P less than 0.001) and moderate AIH and light AIH (P less than 0.001). There is no statistical significance between both light AIH and severe AIH (P = 0.145 ) and moderate AIH and severe AIH (P = 0.084). As a negative regulatory factor, the expression of SIRPalpha1 in hepatic sinusoid Kupffer cells is some associated with the extent of AIH.

Immune cell-specific delivery of beta-glucan-coated iron oxide nanoparticles for diagnosing liver metastasis by MR imaging

Glucans are reported to elicit immune responses through activation of macrophages by a specific interaction of β-glucan with an immune cell-specific (1,3)-β-d-glucan receptor or Dectin-1 receptor. In this study, superparamagnetic iron oxide nanoparticles (SPIONs) were coated with β-glucan in order to target the immune cells residing in the metastatic liver as an aid for discriminating metastasized tumor regions from normal hepatic parenchymal tissue. The morphology of prepared β-glucan-coated SPIONs (Glu-SPIONs) was characterized with dynamic light scattering (DLS) and transmission electron microscopy (TEM). The cytotoxicity of Glu-SPIONs was analyzed and compared to that of dextran- and PVA-coated SPIONs. The uptake of Glu-SPIONs by peritoneal macrophages was also confirmed with Prussian blue staining and MRI phantom tube imaging. The in vivo uptake of Glu-SPIONs in liver and lymph nodes in a metastatic mouse liver model was tracked by MR imaging after the systemic injection. The Glu-SPIONs predominantly accumulated in the macrophages surrounding the metastatic regions of the liver thereby indicating the distribution of tumor cells in the liver. MR imaging of the Glu-SPIONs clearly revealed macro- or micro-metastasized tumor regions throughout the liver, due to the preferential uptake of Glu-SPIONs into macrophages, not tumor cells. The Glu-SPION-accumulating regions were further confirmed with H&E and Prussian blue stainings after tissue sectioning. Based on our study, we propose that Glu-SPIONs can be successfully applied for diagnosing hepatic metastasis.

The expression and significance of the correlation factor of unfolded protein response: Bip in hepatoma

Objective To detect the express of immunoglobulin-binding protein (Bip) mRNA in the tissue of hepatoma;demonstrating that it exist activation of unfold protein response (UPR) in process of growth, and the expression of UPR was higher than normal tissue. Methods To study the mRNA of Bip in 42 case example of hepatome, 15 case paired adjacent hepatic tissues to carcinoma and 15 case normal hepatic tissues by RT-PCR. To qualitation check the expression of Bip in the level of protein by Western blotting. Results The express of Bip mRNA in hepatoma tissue、adjacent hepatic tissue to carcinoma and normai hepatic tissue respectively was 0. 4772 ± 0. 0401、0. 4332 ± 0. 0488、0. 4301 ± 0. 0398; the express of Bip mRNA was higher in hepatoma than others (P ＜ 0. 05 ). The experiment indicate: the Bip express in the hepatoma tissue and normal hepatic tissue, but it was higher in hepatoma tissue than in the normal hepatic tissue. Conclusion It exist activation of UPR in process of growth;The express of Bip protern and its mRNA was accordant, it was higher in hepatoma tissue than in normal hepatic tissue. The changes of its gene and protern is possible to have the relationship with the carcinogenesis, growth and transfer of hepatoma. And it may have significant effect in molecule mechanism of hepatoma. It also descripte that unfold protein response may have very important role in the pocess of carcinogenesis. Key words: Hepatoma; Hepatic tissue; Bip; RT-PCR; Western blotting

Cytoplasmic Staining of TTF-1 Antibody in the Diagnosis of Hepatocellular Carcinoma: Study on 299 Cases Using Tissue Microarray

The diagnosis of hepatocellular carcinoma (HCC) remains obscure in some complicated cases. Thyroid transcription factor-1 (TTF-1) was presumed to be helpful in the diagnosis of HCC. This paper aims to study the utility of TTF-1 in the diagnosis and differential diagnosis of HCC and to assign clinicopathological correlations. Immunohistochemical detection of TTF-1 is performed with tissue microarray which contains a total of 781 cases including 299 hepatocellular carcinomas (HCCs), 31 intrahepaticcholangiocarcinomas (ICCs), 86 metastatic adenocarcinomas (MACs), 5 metastatic hepatocellular carcinomas (mHCCs), 9 hepatoblastomas (HBs), 40 renal clear cell carcinomas (RCCs), 215 adjacent nonneoplastic hepatic tissue, and 96 normal hepatic tissue. The result shows that TTF-1 is positive in 55.2% (165/299) HCCs, 6.4% (2/31) ICCs, 3.5% (3/86) MACs, 60% (3/5) mHCCs, 33.3% (3/9) HBs, 0% (0/40) RCCs, 99.0% (213/215) adjacent nonneoplastic hepatic tissue, and 95.8% (93/96) normal hepatic tissue. Expression of TTF-1 is correlated with differentiation of HCC, but not with patients' age, gender, serum AFP, tumor size, number of tumor nodules, and tumor thrombus in vein or bile duct. These data suggest that TTF-1 antibody is sensitive and specific in HCC and might serve as a candidate for the diagnosis and differential diagnosis of HCC.

Tissue factor pathway inhibitor-2 inhibits the growth and invasion of hepatocellular carcinoma cells and is inactivated in human hepatocellular carcinoma

Human tissue factor pathway inhibitor-2 (TFPI-2) is an extracellular matrix-associated Kunitz-type serine proteinase inhibitor that inhibits the plasmin- and trypsin-mediated activation of matrix metalloproteinases and inhibits tumor progression, invasion and metastasis. Previous studies have shown that TFPI-2 is downregulated in the progression of various tumors. The purpose of this study was to investigate the expression and function of TFPI-2 in hepatocellular carcinoma (HCC). In situ hybridization was used to detect human TFPI-2 mRNA and immunohistochemistry was performed to examine the role of TFPI-2 expression in hepatocarcinoma tissues. Cell proliferation was assessed using MTT assay. Insitu hybridization and immunohistochemical analyses revealed that the expression of TFPI-2 in hepatocarcinoma tissues was markedly lower than that in tumor-adjacent normal hepatic tissues. Restored expression of TFPI-2 in HepG(2) cells inhibits cell proliferation and invasion. Taken together, the results suggest that TFPI-2 has a tumor-suppression action and its inactivation may contribute to HCC.

Comparison of the Anti-tumor Effects of Two Platinum Agents (Miriplatin and Fine-Powder Cisplatin)

This study was designed to evaluate the anti-tumor effects of miriplatin-lipidol and fine-powder cisplatin-lipiodol suspensions. Assessment of the cytotoxicity of two drugs was performed: a soluble derivative of miriplatin (DPC) and fine-powder cisplatin. We randomly divided 15 rabbits with transplanted VX2 liver tumors into three equal groups. They were infused via the proper hepatic artery with a miriplatin-lipiodol suspension (ML), a fine-powder cisplatin-lipiodol suspension (CL), or saline (control) and the tumor growth rate was determined on MR images acquired before and 7 days after treatment. The concentration of platinum (PCs) in blood was assayed immediately, and 10, 30, and 60 min, and 24 h and 7 days after drug administration. Its concentration in tumor and surrounding normal liver tissues was determined at 7 days postadministration. At high concentrations, fine-powder cisplatin exhibited stronger cytotoxicity than DPC. At low concentrations, both agents manifested weak cytotoxicity. While there was no difference between the tumor growth rate of the ML and the CL groups, the difference between the controls and ML- and CL-treated rabbits was significant. The blood PCs peaked at 10 min and then gradually decreased over time. On the other hand, no platinum was detected at any point after the administration of ML. There was no difference between the ML and CL groups in the PCs in tumor tissues; however, in normal hepatic tissue, the PCs were higher in ML- than CL-treated rabbits. We confirmed the anti-tumor effect of ML and CL. There was no significant difference between the anti-tumor effect of ML and CL at 7 days postadministration.

Preliminary screening and analysis on mRNA relating to liver cancer in HBV-associated hepatocellular carcinoma

Objective To explore mRNA expression profile variation of hepatitis B virus ( HBV)related hepatocellular carcinoma and analyze bioinformazics. Methods Long-chain RNA chip was used to inspect mRNA expression profile in HBV-related hepatocellular carcinoma and normal hepatic tissue and make GO analysis for function and cellular localization of genes with differential expression. Results By comparing mRNA expression level of liver cancer with normal control group, 991 mRNA which had more than 2 times variation and significant difference (P＜0.05 ) by statistical analysis were regarded as differential expression mRNA, accounting for 5.7% of all genes. 733 mRNA were increased by more than 2times and 258 decreased by more than 2 times; 94 increased by more than 5 times, 39 decreased by more than 5 times; 18 increased by more than 10 times, 11 decreased by more than 10 times. GO analysis revealed that differential expression mRNA involved in significant biological regulatory function, and some genes may be particular to HBV-related liver cancer. Conclusion The mRNA expression profile in HBVrelated liver cancer changes significantly, and the different genes participate in significant biological regulation process of cells and maybe participate in significant regulation process of liver cancer; HBV-related liver cancer' s expression profile has certain characteristics and something in common with liver cancers relating to other pathogenesis. Key words: Carcinoma,hepatocellular; Gene microarray; Hepatitis B virus

SLC25A13 gene mutations in Taiwanese patients with non-viralhepatocellular carcinoma

Mutations of the SLC25A13 gene, which encodes citrin, result in adult-onset type II citrullinemia (CTLN2). Because CTLN2 has been associated with hepatocellular carcinoma (HCC) and may be involved in hepatocarcinogenesis, the objective of this study was to assess the frequency of SLC25A13 mutations in patients with non-viral HCC. A retrospective review of 154 patients with HCC, who underwent total tumor resection from July 1998 to August 2005, was conducted. After exclusion of 137 patients infected with hepatitis B and/or C viruses, 17 patients were analyzed. Genomic DNA from stored tumor and normal hepatic samples was analyzed for the SLC25A13 gene mutation. In addition, the clinicopathological and histopathological features of patients with and without the SLC25A13 gene mutation were compared. The SLC25A13 mutation was observed in two patients (12%), and the carrier rate was approximately 1 in 8 patients. The IVS6 + 5G>A mutation was heterozygous in both normal hepatic and tumor tissues for case 1. On the other hand, the c.851del4 mutation was heterozygous in normal tissue but homozygous in tumor tissue for case 2. No significant differences in patient characteristics were observed. Further analyses of patients with SLC25A13 gene mutations may elucidate the relationship between the citrin gene and susceptibility of HCC.

Imaging of blood flow using hyperpolarized [13C]Urea in preclinical cancer models

To demonstrate dynamic imaging of a diffusible perfusion tracer, hyperpolarized [(13)C]urea, for regional measurement of blood flow in preclinical cancer models. A pulse sequence using balanced steady state free precession (bSSFP) was developed, with progressively increasing flip angles for efficient sampling of the hyperpolarized magnetization. This allowed temporal and volumetric imaging of the [(13)C]urea signal. Regional signal dynamics were quantified for kidneys and liver, and estimates of relative blood flows were derived from the data. Detailed perfusion simulations were performed to validate the methodology. Significant differences were observed in the signal patterns between normal and cancerous murine hepatic tissues. In particular, a 19% reduction in mean blood flow was observed in tumors, with 26% elevation in the tumor rim. The blood flow maps were also compared with metabolic imaging results with hyperpolarized [1-(13)C]pyruvate. Regional assessment of perfusion is possible by imaging of hyperpolarized [(13)C]urea, which is significant for the imaging of cancer.

Preparation of EUS-guided radiofrequency probe and test of its functions

Objective To prepare a radiofrequeney （RF） ablation probe guided by endoscopic ultrasonography （EUS） and to evaluate its efficacy and safety. Methods A modified 19-gange needle （ Olympus） was used, which was electrically insulated with shrink tubing, with the tip of lcm naked. The insulation was tested, and the current signal was stable. Three rabbits were anesthetized with 8% chloral hydrate （3 ml/kg） intraperitoneally, fixed on the dissecting table. A standard neutral pad was applied to the thigh of the rabbit to complete the electrical circuit, both the pad and the needle electrode connected with RF generator. After anesthesia was ready, the needle was advanced through the gastric wall into the liver. Ablations were performed three times in the same zone. Extent and tissue lesion were measured after ablation. Results Ablation could be successfully performed by the needle electrode, with brown lesions surrounded by normal hepatic tissue. The mean diameter of the ablated zone in the liver was 1.0 cm × 1.2 cm. After ablation, specimens along the passage were subjected to NADH staining, and no lesions were found. HE and NADH staining showed no viable cells in the central ablation area. Conclusion With the advance of the technology and selection of materials, an EUS-guided needle electrode can be made. This preliminary animal trial dem- onstrates that radiofrequency ablation can be performed effectively and safely by using this EUS-guided needle electrode. Key words: Endoscopic ultrasonography ; Electrodes ; Catheter ablation, radiofrequency

Expression of HIWI in Human Hepatocellular Carcinoma

In this study, the investigation of the expression of HIWI and its protein in hepatocellular carcinoma (HCC) was performed, and the relationships between HIWI expression and the location of HCC metastases were analyzed. Sets of fresh HCC and matched adjacent normal hepatic tissue and paraffin-embedded tissue slides were provided by the hospital hepatology and pathology departments. RT-PCR, Western blot, and immunohistochemistry were performed to detect HIWI mRNA and protein. Correlations between HIWI expression and patient's age, sex, type of tumor, and metastasis location were recorded. HIWI mRNA and protein levels were significantly higher in HCC tissues than in adjacent normal hepatic tissue (P<0.05). Immunohistochemistry showed positive staining for HIWI in cell cytoplasm; however, the number of HIWI-positive cells in HCC tissue (65.2%; 60/92) was significantly higher than in adjacent normal hepatic tissue (27.2%; 25/92) (P<0.05). HIWI expression was not correlated with patients' age, gender, tumors' size, and location but correlated with metastasis involving lymph nodes and other remote organs (P<0.05). HIWI expression is significantly higher in HCC tissue than in adjacent normal hepatic tissue. The results of this study suggest that HIWI may have a crucial role in HCC carcinogenesis and could serve as a potential biomarker or treatment target for HCC.

Hepatocellular carcinoma with marginal superparamagnetic iron oxide uptake on T2*-weighted magnetic resonance imaging: Histopathologic correlation

To evaluate the characteristics of hepatocellular carcinomas (HCCs) with marginal superparamagnetic iron oxide (SPIO) uptake on T2*-weighted MRI. The study group consisted of 73 patients with 83 surgically resected HCCs. Preoperative SPIO-enhanced MRI studies were retrospectively reviewed. Marginal SPIO uptake was considered positive if a rim-like or band-like low intensity area was present on SPIO-enhanced T2*-weighted images. The prevalence of marginal SPIO uptake was evaluated. Pathological specimens with hematoxylin and eosin staining and immunohistochemical staining of CD68 were reviewed in HCCs with marginal SPIO uptake and 33 HCCs without marginal SPIO uptake (control group). Ten of 83 (12%) HCCs showed marginal SPIO uptake. All HCCs were hypervascular, and only one nodule showed a nodule-in-nodule appearance on imaging findings. The pathology specimens suggested possible causes of marginal SPIO uptake, including marginal macrophage infiltration in moderately or poorly differentiated HCC (n=4), residual normal hepatic tissue at the marginal area of confluent multinodular or single nodular with extranodular growth type HCC (n=3), and a well-differentiated HCC component in nodule-in-nodule type HCC (n=3). Marginal macrophage infiltration was not seen in the control group. SPIO-enhanced MRI may be able to demonstrate marginal macrophage infiltration in HCC.

Down-regulation of hemojuvelin prevents inhibitory effects of bone morphogenetic proteins on iron metabolism in hepatocellular carcinoma

Recently, we revealed that bone morphogenetic protein (BMP) 4 is increased in hepatocellular carcinoma (HCC), and latest reports describe BMPs, in particular BMP6, as important regulators of hepcidin expression in iron homeostasis. The aim of this study was to unravel why enhanced BMP expression in HCC does not lead to severe changes in iron metabolism. Methods and Results: Initial analysis confirmed and newly revealed, respectively, enhanced mRNA and protein expression of both BMP4 and BMP6 in human HCC cell lines and tissues compared to primary human hepatocytes (PHH) and normal liver tissues. However and surprisingly, hepcidin expression was reduced in HCC cell lines and tissues. Analysis of BMP6 receptor expression revealed loss of BMP6 specific receptor subunit in HCC. To identify a possible regulatory mechanism causing the lack of response towards BMP4 we analyzed the expression of hemojuvelin (HJV), which is involved in iron metabolism acting as a BMP co-receptor. HJV expression was markedly decreased in HCC cell lines and tissues, and moreover, also in surrounding non-tumorous liver tissue. In line with this, we found reduced hepatic HJV expression in an experimental murine model bearing BMP expressing tumors. HJV promoter analysis revealed potential HNF-1α and snail binding sites, but functional analysis ruled out that these transcriptional regulators or promoter methylation are the cause of HJV down-regulation in HCC. However, we identified AU-rich elements in the HJV 3’-untranslated region. We revealed significantly faster decay of HJV mRNA in HCC cells as compared to PHH indicating decreased mRNA-stability as the reason for loss of HJV expression in HCC. Conclusion: Hepatic tumors decrease hemojuvelin expression, and herewith, inhibit inhibitory effects of BMPs on iron levels. Thus, regulation of iron homeostasis in HCC differs significantly from normal hepatic tissue.

Contrast-enhanced ultrasound in planning thermal ablation of liver metastases: Should the hypervascular halo be included in the ablation volume?

Introduction Liver metastases often exhibit a hypervascular halo during the arterial phase of contrast-enhanced ultrasonography (CEUS). This finding has no correlates on baseline gray-scale imaging, and it has never been characterized. The aim of this study was to identify the features of this halo and determine whether it should be included in the ablation volume during thermal ablation procedures. Materials and methods We prospectively enrolled 25 patients referred to our department for thermal ablation of liver metastases. Before treatment all patients underwent CEUS, and the maximum diameter of the metastatic lesion was measured before administration of the ultrasound contrast agent and during the arterial and portal venous phases of the contrast contrast-enhanced study. Maximum diameters in the different vascular phases were compared with the Turkey–Kramer test. Two biopsies were obtained from each lesion with a 21-gauge needle: 1) one from the center of the metastasis to confirm the diagnosis and 2) one from the hypervascular peripheral halo identified in the arterial phase at CEUS. Results The mean (±standard deviation) maximum lesion diameter was 2.67 ± 1.2 cm before contrast agent injection, 3.50 ± 1.4 cm during the arterial phase, and 2.71 ± 1.2 cm during the venous phase. The difference between maximum diameters measured before contrast enhancement and in the arterial phase was highly significant (mean: 0.84 ± 0.45 cm, p < 0.0001). Histological examination of halo specimens revealed inflammatory infiltrates with no evidence of tumor infiltration in 24/25 (96%) cases and normal hepatic parenchymal tissue in the 25th specimen. Discussion The hypervascular halo surrounding liver metastases during the arterial phase of CEUS represents a chronic inflammatory infiltrate, not tumor infiltration. However, since chronic inflammation appears to promote neovascularization and the production of tumoral growth factors, it seems wise to include the hypervascular halo in the intended-to-treat volume when planning the ablation procedure.

Investigation of serum concentrations and immunohistochemical localization of α1-acid glycoprotein in tumor dogs

The purpose of this study was to measure the dynamics of serum α1-acid glycoprotein (AGP) concentration in dogs with various tumors, and to investigate the localization of AGP in some tissues using immunohistochemical staining. Sera were obtained from 171 dogs bearing tumors of various types. Serum AGP concentration was measured by single radial immunodiffusion. Tumors occurring in the liver and spleen were also investigated immunohistochemically using anti-canine AGP antibody. Mean serum AGP levels were 749 ± 602 mg/L in dogs with carcinoma (n = 39), 1,014 ± 971 mg/L with sarcoma (n = 18), and 887 ± 935 mg/L with round cell tumors (n = 46), all significantly higher than serum AGP level in healthy dogs (n = 137, 364 ± 106 mg/L). Mean serum AGP levels were significantly higher than in healthy dogs in complex mammary gland carcinoma (n = 5, 876 ± 721 mg/L), malignant melanoma (n = 7, 1,010 ± 821 mg/L), and hepatocellular carcinoma (n = 5, 936 ± 741 mg/L) among carcinomas, hemangiosarcoma (n = 5, 1,740 ± 1,323 mg/L) among sarcomas, and lymphoma (n = 19, 1,072 ± 965 mg/L) and histiocytic tumor (n = 6, 1,800 ± 1,387 mg/L) among round cell tumors. In an immunohistochemical investigation of AGP localization, both weak and strong staining for anti-AGP antibody were seen in hepatic tissue in dogs with primary non-tumorous lesions originating in the spleen (hematoma) and elevated serum AGP, but all tumor tissue in the spleen was negative. Among dogs with primary tumor lesions of the spleen (hemangiosarcoma) and elevated serum AGP levels, both weak, moderate and strong staining for anti-AGP antibody were seen in hepatic tissue, while strong positive staining was apparent in all tumorous tissue from the spleen. In primary tumor lesions in the liver (hepatocellular carcinoma), both moderate and strong staining for anti-AGP antibody were seen in normal hepatic tissue, and both weak, moderate and strong staining were seen in tumor tissues of the liver. AGP levels thus appear to be elevated in dogs with carcinomas, sarcomas, and round cell tumors. With some of these malignant tumors, localization of AGP in tumor tissue was seen.

Electrical impedance characterization of normal and cancerous human hepatic tissue

The four-electrode method was used to measure the ex vivo complex electrical impedance of tissues from 14 hepatic tumors and the surrounding normal liver from six patients. Measurements were done in the frequency range 1–400 kHz. It was found that the conductivity of the tumor tissue was much higher than that of the normal liver tissue in this frequency range (from 0.14 ± 0.06 S m−1 versus 0.03 ± 0.01 S m−1 at 1 kHz to 0.25 ± 0.06 S m−1 versus 0.15 ± 0.03 S m−1 at 400 kHz). The Cole–Cole models were estimated from the experimental data and the four parameters (ρ0, ρ∞, α, fc) were obtained using a least-squares fit algorithm. The Cole–Cole parameters for the cancerous and normal liver are 9 ± 4 Ω m−1, 2.2 ± 0.7 Ω m−1, 0.5 ± 0.2, 140 ± 103 kHz and 50 ± 28 Ω m−1, 3.2 ± 0.6 Ω m−1, 0.64 ± 0.04, 10 ± 7 kHz, respectively. These data can contribute to developing bioelectric applications for tissue diagnostics and in tissue treatment planning with electrical fields such as radiofrequency tissue ablation, electrochemotherapy and gene therapy with reversible electroporation, nanoscale pulsing and irreversible electroporation.

Up-regulation of Tiam1 and Rac1 Correlates with Poor Prognosis in Hepatocellular Carcinoma

T-cell lymphoma invasion and metastasis 1 (Tiam1) specifically activates Rho-like GTPases (e.g. Rac1) and Tiam1-Rac1 pathway affects the migration and invasion of many tumors, such as nasopharyngeal carcinoma, breast cancer and retinoblastoma. However, no studies have yet comprehensively examined the involvement of Tiam1-Rac1 pathway in hepatocellular carcinoma. In this study, we examined the relationship of the up-regulation of Tiam1 and Rac1 with clinicopathological features in patients with hepatocellular carcinoma. Expression of Tiam1 and Rac1 was assessed in 242 hepatocellular carcinoma tissues and their adjacent normal hepatic tissues by performing immunohistochemistry and was gauged regarding stage, grade and survival. Immunohistochemistry showed that patients with a high clinical stage hepatocellular carcinoma (III-IV) and α-fetoprotein levels had a higher tendency to express Tiam1 and Rac1 on tumor cells than the patients with low pathologic grade hepatocellular carcinoma (I-II) (P = 0.008 and 0.01, respectively) and low α-fetoprotein levels (P = 0.006 and 0.002, respectively). In addition, Tiam1 and Rac1 up-regulation was also significantly associated with vascular invasion status (both P = 0.02), intrahepatic metastasis status (P = 0.009 and 0.01, respectively) and histological differentiation (P = 0.008 and 0.009, respectively) of patients with hepatocellular carcinoma. Moreover, post-operative survival analysis indicated that hepatocellular carcinoma patients with strong Tiam1 (P = 0.01) and Rac1 (P = 0.02) expression had shorter disease-specific survival than those with weak expression. Multivariate analysis also showed that Tiam1 and Rac1 overexpression could be two predictors of poor prognosis (P = 0.02 and 0.03, respectively). The current study demonstrated for the first time that the Tiam1-Rac1 pathway may play a critical role in tumor progression of hepatocellular carcinoma. The expression of Tiam1 and Rac1 can be considered as the two useful indicators for predicting the prognosis of hepatocellular carcinoma.

The expression of integrin beta 1 in normal hepatic tissues, hepatic cirrhosis tissues and hepatocellular carcinoma

To investigate the expression of integrin beta 1 in hepatic cirrhosis (HC) and hepatocellular carcinoma (HCC). The expression of integrin beta 1 in HCC, HC and normal liver tissues was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and laser scanning confocal microscopy (LSCM). The association between the integrin beta 1 expression and clinical pathological features were analyzed. (1) The levels of integrin beta 1 mRNA and protein in the HCC (1.30+/-0.24, 90.50+/-33.50) and HC (1.58+/-0.31, 123.10+/-38.90) were much higher than that in the normal hepatic tissue (0.37+/-0.08, 11.90+/-6.00) (P less than 0.05). (2) The expression of integrin beta 1 was associated with HC (r = 0.692), Edmondson pathologic grade (F = 13.618), encapsulation (F = 17.857) and metastasis (F = 38.857) (P less than 0.01). Integrin beta 1 may play an important role in the development of hepatic fibrosis, hepatic cirrhosis and hepatocellular carcinoma.