Five-year survival of patients with hepatocellular carcinoma treated with laparoscopic microwave coagulation therapy.

BACKGROUND AND STUDY AIM: Hepatocellular carcinomas (HCCs) that are located near the liver surface are difficult to treat with percutaneous locoregional therapies, so we have performed laparoscopic microwave coagulation therapy (LMCT) for these HCCs. We assessed the long-term survival of patients with HCCs treated with LMCT, the factors related to their survival, and the rates and causes of local and distant recurrences. PATIENTS AND METHODS: Participants were 68 patients with HCC treated in the past 8 years with LMCT under local or general anesthesia. LMCT was done using microwave electrodes with tips ranging from 15 to 45 mm in length; the effectiveness of LMCT was confirmed using contrast-enhanced computed tomography (CT) within 2 weeks of the LMCT procedure while patients were still in hospital, and within 1-3 months after the procedure in an outpatient setting; and the follow-up study was performed periodically by CT, ultrasonography, or magnetic resonance imaging (MRI) in addition to estimation of alpha fetoprotein. Factors contributing to survival were analyzed statistically. RESULTS: The mean lengths of the major and minor axes of the 71 HCC nodules in 68 patients were 20 mm and 18 mm, respectively, and the mean lengths of the major and minor axes of the coagulated areas were 43 mm and 29 mm, respectively. At dynamic CT after the LMCT procedure, treatment in 62 of the 68 patients (91%) was judged to have been completely effective; the remaining six patients underwent additional therapy while still in hospital. Eight of the 68 patients (12%) had local recurrences, 39 of them (57%) had distant recurrences, and 21 of them (31%) had no recurrence up to December 31, 2003. A total of 14 patients (21%) died during the 16-56 months after LMCT. When the survival rate was assessed for all patients treated with LMCT, 1-year survival was 97 %, 3-year survival was 81%, and 5-year survival was 43%. Whether the therapy was for primary or secondary HCC strongly influenced survival. CONCLUSIONS: LMCT is a useful modality in clinical practice for treatment of HCC nodules located near the liver surface, and it can be safely performed, in its entirety, under direct visual guidance.

Unusual nature of ferromagnetism coexisting with superconductivity in UGe2.

We report the discovery of a jump in the magnetization of a macroscopic single crystalline sample of UGe2 that shows coexistence of ferromagnetism and superconductivity. In particular, we observe that the jump occurs at regular intervals of field and only at very low temperatures. This novel feature implies that the magnetic field induces a sudden change of the direction of the magnetization between two equivalent easy axes of magnetization even in a macroscopic sample. We ascribe it to a field-tuned resonant tunneling between quantum spin states, and we propose that the size of a magnetic domain is smaller than a superconducting coherence length.

Cellular prion protein expression in non-ciliated epithelial cells (Clara cells) of proliferating bronchioles during bleomycin-induced pulmonary fibrosis in hamster.

The cellular isoform of prion protein (PrPc) is a membrane glycoprotein with unknown roles. This study aimed to immunohistochemically demonstrate PrPc expression in normal and fibrotic hamster lungs. In untreated lungs, Clara cells, which were preferentially distributed in small bronchioles and characterized by globule granules, were positively stained. Reaction products were diffuse in the cytoplasm outside the granules and nuclei. In bleomycin-induced pulmonary fibrosis, bronchiolar proliferation was induced in the thickened fibrous tissue around terminal bronchioles. This event was characterized by accumulation of ductular structures which were predominantly lined by PrPc positive cells representative of Clara cells. Furthermore, PrPc positive cells were occasionally populated in the epithelium of alveolar ducts and reepithelialized alveoli. These findings suggest that Clara cells might undergo proliferation and migration into acini from terminal bronchioles. The fibrous tissue contained many alpha-smooth muscle actin positive myofibroblasts. The present study indicates that PrPc is expressed in Clara cells in normal and fibrotic hamster lungs and suggests that Clara cells may proliferate as pulmonary stem cells and repair the damaged epithelium after alveolar injuries.

Clinicopathological characterization of prion: a novel marker of activated human hepatic stellate cells.

BACKGROUND/AIMS: We recently demonstrated prion as a new marker for hepatic stellate cell activation in rats. Here, we have examined prion expression in normal and diseased human livers. METHODS: Prion expression was examined at protein level by immunohistochemistry and at mRNA level by in situ hybridization. RESULTS: While normal livers were negative for prion, all liver specimens but one from patients with chronic liver disease were positively stained. In chronic hepatitis, prion protein expression was found not only in the sinusoidal lining cells within the lobules but also in mesenchymal cells in expanded portal tracts. In alcoholic liver disease, prion-positive cells were found mainly in the areas of alcoholic hepatitis. Immunoelectronmicroscopy revealed that prion-positive cells were activated stellate cells. In situ hybridization demonstrated that the distribution of prion mRNA is similar to that of prion protein. In chronic hepatitis, the number of prion-positive cells correlated with the grade of activity but not with the stage of fibrosis. In alcoholic liver disease, levels of prion protein expression were significantly increased in the presence of alcoholic hepatitis. CONCLUSION: Prion as a novel marker of activated stellate cells correlates well with disease activity in human chronic liver diseases.

Laparoscopic microwave coagulation therapy for hepatocellular carcinoma.

BACKGROUND AND STUDY AIMS: Several different effective forms of treatment are available, singly or in combination, for patients with hepatocellular carcinoma (HCC). These include surgical resection, transcatheter arterial embolization, percutaneous ethanol injection, and percutaneous microwave coagulation therapy. In this study, we carried out laparoscopic microwave coagulation therapy (LMCT), using laparoscopic microwave electrodes to treat HCC. PATIENTS AND METHODS: Under local anesthesia, 24 patients with HCCs located on or near the liver surface underwent LMCT under direct laparoscopic vision, with ultrasound guidance. LMCT was performed using microwave electrodes with tips ranging from 15-45 mm in length, and the effectiveness of the treatment was confirmed using contrast-enhanced computed tomography (CT) within two weeks of the LMCT procedure. RESULTS: The mean longest axis of the 26 HCC nodules in 24 patients was 20 mm, and that of the coagulated areas including the nodules was 40 mm, with additional therapy being required in two patients. Complete efficacy of the treatment was observed in 21 patients (87.5%), but local recurrences were seen in three of them one year after LMCT. The three-year survival rate was 92%, but the number of patients included in the study was small. Hemostasis was complete, but mild pneumothorax occurred in three patients. CONCLUSIONS: LMCT under local anesthesia is a minimally invasive and effective therapy when carried out on a single occasion to treat HCCs located near the liver surface, and it can be safely performed under direct visual guidance.

Establishment and characteristics of human hepatocellular carcinoma cells with metastasis to lymph nodes.

BACKGROUND/AIMS: Although the mechanism of cancer metastasis has been gradually elucidated, less is known concerning the characteristics of human hepatocellular carcinomas (HCCs) with metastatic potential. We examined the expression of molecules that mediate cell-cell or cell-substrate interaction, nm23-H1 expression, and ultrastructural features of several human HCC cell lines. METHODOLOGY: Expression of E-cadherin, integrin (alpha3beta1), intracellular adhesion molecule-1 (ICAM-1), and nm23-H1 protein was analyzed by immunocytochemistry or Western blotting, and ultrastructural features were further studied by electron microscopy in 4 human HCC cell lines, PLC/PRF/5, HuH-7, OCUH-16, and Nuk-1 which were originally established from metastatic cells in lymph nodes at our institute. RESULTS: Neither E-cadherin, integrin, nor ICAM-1 was immunocytochemically detected in any of the 4 cell lines. Expression of nm23-H1 protein was weakly detected in OCUH-16, Nuk-1, and Huh-7 cells by Western blotting, but was clearly detected in PLC/PRF/5 cells by Western blotting. Ultrastructurally, metastatic Nuk-1 cells exhibited the intracytoplasmic canaliculus-like structures found in fibrolamellar carcinoma and the intracytoplasmic glandular lumina found in bile-duct carcinoma, while the other 3 cell lines did not. In addition, Nuk-1 cells expressed neither cytokeratin 8 nor cytokeratin 19. CONCLUSIONS: Nuk-1 cells, which are human HCC cells with metastasis to lymph nodes, alone exhibited intracytoplasmic canaliculus-like structures and glandular lumina, as well as a marked reduction of nm23-H1 protein, but did not express E-cadherin, integrin, or ICAM-1. Formation of both intracytoplasmic canaliculus-like structures and intracytoplasmic glandular lumina is one of the characteristics that may be involved in metastasis of HCC cells to lymph nodes, as is reduction of nm23-H1 protein.

In vitro migratory potential of rat quiescent hepatic stellate cells and its augmentation by cell activation.

In liver injury, hepatic stellate cells are considered to depart from the sinusoidal wall and accumulate in the necrotic lesion through migration and proliferation. In this study, we investigated the migratory capacity of quiescent stellate cells in vitro and analyzed the relationship with proliferative response. Freshly isolated stellate cells that were seeded in the upper chamber of Cell Culture Insert (Becton Dickenson, Franklin Lakes, NJ) started to migrate to the lower chamber at 1 day and increased in migration index to 19% at 2 days. Cells in the lower chamber were stretched in shape with many lipid droplets and showed quiescent properties, i.e., negative expression of alpha-smooth muscle actin (alpha-SMA) or platelet-derived growth factor receptor-beta (PDGFR-beta). Migratory capacity in quiescent cells was also shown in the Matrigel-coated insert. Matrix metalloproteinase-2 (MMP-2) messenger RNA expression was low just after isolation, but was enhanced as migration became prominent. Migrating cells further showed higher proliferative activity than resting ones. The presence of PDGF/BB and Kupffer cells accelerated stellate cell migration by the chemotactic mechanism and concurrently augmented proliferation, whereas that of dexamethasone and interferon-gamma (IFN-gamma) attenuated migration as a result of general suppression effects. Compared with quiescent ones, alpha-SMA and PDGFR-beta-positive activated stellate cells obtained by 14-day culture exhibited more rapid and prominent migration, being regulated by mediators in a similar manner as described previously. These data indicate that quiescent stellate cells undergo migration, which is linked to proliferation and enhanced by PDGF/BB and Kupffer cells, suggesting the involvement of this function in the initial phase of development of postnecrotic fibrosis.

Expression of cellular prion protein in activated hepatic stellate cells.

Suppression subtractive hybridization was used to clone genes associated with the activation of hepatic stellate cells and 13 genes were found to be dominantly expressed in activated stellate cells. Among them, one was identical to the 421-837th base pairs of cDNA sequence reported for rat prion-related protein (PrP). In cultured stellate cells, PrP mRNA expression increased in a time-dependent manner in parallel with smooth muscle (SM) alpha-actin mRNA expression. In situ hybridization demonstrated that PrP mRNA was localized in and around the fibrous septa of carbon tetrachloride (CCl4)-treated liver. Cellular PrP (PrPc) was produced by culture-activated stellate cells, and immunohistochemically detected in the fibrous septa of CCl4-damaged liver and sinusoidal linings of common bile duct-ligated liver, consistent with the localization of SM alpha-actin. Immunoelectron microscopy revealed that PrPc resided on the plasma membrane of stellate cells. These results indicate that PrP expression is closely related to stellate cell activation associated with fibrogenic stimuli.

Ultrastructural study of a perineurioma.

A case of soft tissue tumor in the left brachialis muscle of a 49-year-old Japanese female patient was studied by electron microscopy. The tumor was diagnosed as intramuscular myxoma by light microscopy, but electron microscopic observation revealed that the tumor almost entirely consisted of cells similar to normal perineurial cells. The tumor cells possessed long, slender cytoplasmic processes covered by well-developed but discontinuous basal laminae, clusters of pinocytotic vesicles, and infrequent intercellular junctions. Perineurial cells have also been observed in other peripheral nerve lesions: neurofibromas, nerve sheath myxomas, and localized hypertrophic neuropathies. However, the term "perineurioma" or "perineurial cell tumor" should be reserved for discrete tumorous masses that are almost entirely composed of perineurial cells without evidence of residual axons, Schwann cells, fibroblasts, or tactile corpusclelike structures. Perineurioma may represent a third category of peripheral nerve sheath tumors, ultrastructurally distinct from schwannomas and neurofibromas.

A 20,000-dalton casein fragment in human milk.

A new peptide of 20,000 daltons was found in human milk as a constituent of the casein micelle. Enzymic digestion with plasmin or trypsin revealed that the peptide was identical with a degradation product of human beta-casein. The amino acid composition of the degradation product and the previously reported sequence in the N-terminal region of human beta-casein suggested that the peptide was a fragment of beta-casein lacking the C-terminal region. The thermal sensitivity of this peptide was higher than that of beta-casein, but the peptide lost the property of calcium-dependent precipitation, which intact beta-casein possesses.