Establishment of a new human pancreatic cancer cell line, NOR-P1, with high angiogenic activity and metastatic potential.

We present here a new cell line, NOR-P1, established from a metastatic subcutaneous tumor of a patient with pancreatic cancer. The cells show rapid growth in culture with a doubling time of 16 h and high migration activity. Genetic and molecular analyses revealed high telomerase activity and a mutation in the K-ras oncogene. Of particular interest, the cells express markedly elevated mRNA levels of angiogenic factors, vascular endothelial growth factor and platelet-derived growth factor, as well as other tumor growth-related factors. Subcutaneous transplantation of the NOR-P1 cells into nude mice formed solid, hemorrhagic tumors which were histologically diagnosed as adenocarcinoma with dense blood vessels and severe extravasation of blood. Furthermore, when NOR-P1 cell suspension was injected directly into the pancreas of nude mice, the cells grew rapidly to form intra-pancreatic tumors associated with liver metastases and peritoneal dissemination that resulted in cachexia and subsequent death. These properties suggest that NOR-P1 is an aggressive pancreatic cancer cell line with a high metastatic potential and may serve as a useful experimental model for studying tumor angiogenesis and metastasis of pancreatic cancer.

A phase I study of cisplatin i.p. and carboplatin i.v. with G-CSF in patients with ovarian cancer.

We conducted a dose-escalation study with a fixed dose of intraperitoneal cisplatin and G-CSF support of carboplatin using the Calvert formula in epithelial ovarian cancer. Twenty-five patients were entered in this study. On day 1, carboplatin was administered intravenously at target AUCs of 4, 5, 6, and 7. On day 2, cisplatin was given i.p. in 70 mg/m2. G-CSF, 50 microgram/m2, was administered subcutaneously from day 7 to 16. Cycles were scheduled to be delivered every four weeks. A total of 85 cycles were administered. The maximum tolerated dose was AUC 7 mg/ml x min of carboplatin. The overall response rate was 80% (12/15). The combination in this regimen is feasible, and a phase II study of this regimen is warranted.

A combination of cyclosporin-A (CsA) and interferon-gamma (INF-gamma) induces apoptosis in human gastric carcinoma cells.

Modulation of interferon-gamma effect by other drug may enhance its tumor specific activity. The apoptosis inducing effect of interferon-gamma and its modulation by cyclosporin-A or tacrolimus (FK-506) were investigated in in vitro and ex vivo experiments. We found that a combination of cyclosporin-A (CsA) and recombinant interferon-gamma (rIFN-gamma) induced significant apoptosis in all four types of human gastric carcinoma cells tested but not in normal cells such as human peripheral blood mononuclear cells (PBMCs), human omentum-derived mesothelial cells, or human umbilical vein endothelial cells (HUVECs) in vitro. Apoptosis was also induced by a combination of rIFN-gamma with FK-506 but not with rapamycin. Next, the apoptosis-inducing effect of endogenous IFN-gamma combined with cyclosporin-A was examined using clinical samples. A streptococcal preparation, OK-432, was administered intraperitoneally for the management of 12 gastric cancer patients with malignant ascites. None of the gascitic fluids obtained before the OK-432 injection showed detectable IFN-gamma level. The OK-432 injection induced a detectable IFN-gamma production ranging from 6 to 89 pg/mL in ascitic fluids from 9 out of the 12 patients. A combination of CsA with ascitic fluids collected after but not before OK-432 injection induced significant apoptosis in MK-1 cells, a gastric carcinoma cell line. A positive correlation was found between the IFN-gamma level and CsA-induced apoptosis. The CsA-induced apoptosis was also blocked by a specific antibody against human IFN-gamma. These results indicated that both recombinant and endogenous IFN-gamma can induce potent tumor-apoptosis when combined with CsA.

Hepatocyte growth factor production by peripheral blood mononuclear cells of recurrent cancer patients.

The elevation of circulating hepatocyte growth factor (HGF) concentrations is associated with tumor progression and prognosis in cancer patients. We aimed to clarify whether peripheral blood mononuclear cells (PBMC) of cancer patients can produce circulating HGF. In sera and PBMC supematants of 39 cancer patients and 55 control subjects, we measured HGF concentrations by enzyme-linked immunosorbent assay and analyzed HGF expression in PBMC by reverse transcriptase-polymerase chain reaction and Western blotting. Compared with primary cancer patients and control subjects, recurrent cancer patients showed higher serum HGF concentrations (p < 0.05, p < 0.01 respectively), higher HGF concentrations in PBMC supernatants (p < 0.01 for both) and more frequent HGF mRNA expression in PBMC. Additionally, HGF mRNA was induced in PBMC of a control subject cultured with the sera of recurrent cancer patients. These results indicate that PBMC of recurrent cancer patients can produce HGF and that their sera may contain substances that induce HGF mRNA expression.

Elevation of serum hepatocyte growth factor concentration in patients with gastric cancer is mediated by production from tumor tissue.

Hepatocyte growth factor (HGF) is a stromal cell-derived cytokine that can stimulate invasion and metastasis of carcinoma cells. Recent studies have shown that the serum HGF concentration is elevated in patients with gastric cancer and may be a useful disease marker. However, the origin of the elevated serum HGF remains undetermined. We investigated the site of HGF production by analyzing the relationships between the HGF expression in tumor tissues, the serum HGF concentrations and inflammation in patients with gastric cancer. The serum and tissue HGF concentrations were measured by an enzyme-linked immunosorbent assay. The serum HGF concentration was higher than the normal cut-off level (0.57 ng/ml) in 44% of the patients. Surgical removal of the tumor significantly reduced the serum HGF concentration, suggesting that the tumor tissue was responsible for the increase. Western blotting analysis showed that the HGF protein was expressed in 20 out of 22 tumor tissues. The concentration of HGF in the tumor tissue was significantly higher than that in normal gastric mucosa. Significant correlation was found between tissue HGF concentrations and serum HGF concentrations. No significant correlation was found between the serum HGF concentration and white blood cell count or C-reactive protein concentration, indicating that the increase in serum HGF is not due to inflammation related to the tumor. These results suggest that the elevated serum HGF concentration in patients with gastric cancer is mediated by production from the tumor tissue.

A case of multiple sclerosis with multi-ring-like and butterfly-like enhancement on computerized tomography.

We report a case of multiple sclerosis in which CT showed multiple ring-like enhancement and butterfly-like distribution of a low density area with marginal enhancement. The latter finding is found in other demyelinating disorders but is less common in tumors or abscesses. Therefore, it seems to have some diagnostic value in multiple sclerosis.

An iatrogenic foreign body (dental bur) in the maxillary antrum: a report of two cases.

Two cases of foreign bodies of the antrum are reported. One was a turbine bur which presumably entered through an oro-antral fistula after a tooth extraction. The other was also a turbine bur where the mode of entry was not clear (lack of oro-antral fistula), but it presumably entered through the socket of the extracted tooth. The mucosa of the antrum appeared normal in spite of the lengthy presence of the foreign body.

Use of intra-operative ultrasonography to detect a small foreign body in the soft tissue of the upper lip.

Real-time ultrasonography was used to detect a small foreign body (a broken tip of a barbed broach) which had migrated into the upper lip of a 26-year-old female from the bone defect of the upper canine apex during root canal treatment 10 months previously. Intra-operative use of ultrasound was of great-help in clarifying the positional relationship between the foreign body and the surgical instruments used, and lead to a satisfactory result.

Response of healthy individuals to ninjin-yoei-to extract--enhancement of natural killer cell activity.

After administering 15 g/day of Ninjin-Yoei-To (NYT) for one week to healthy people whose NK activity had already been increased by physiological response, a further increase in NK activity was observed after two days. This increased level of NK activity continued during the administration of NYT for a one-week period. The maintaining and reinforcing effect on the immune surveillance system by NYT may be useful for prevention of carcinogenesis.

[Two cases of tracheobronchial disruption due to blunt trauma].

Tracheobronchial disruption by blunt trauma is relative rate, but clinical course is various and sometimes fatal. A 22-year-old man (case 1) was admitted with traffic accident. Chest CT demonstrated pneumomediastinum and left pneumothorax. Bronchoscopy showed tracheal tear, but conservative therapy was selected because the peritracheal sheath remained intact. The second case was 27-year-old man, who was admitted with traffic accident. Pneumomediastinum and left tension pneumothorax were diagnosed, but bronchoscopy could not confirm the tracheobronchial disruption. As bronchial injury, lacerations and contusions of the lingular division were found in the thoracotomy, segmentectomy was carried out. When pneumomediastinum or pneumothorax is recognized in the patient of the blunt trauma, more careful examination of bronchoscopy is necessary to confirm the tracheobronchial disruption.

CCL-1 in the spinal cord contributes to neuropathic pain induced by nerve injury.

Cytokines such as interleukins are known to be involved in the development of neuropathic pain through activation of neuroglia. However, the role of chemokine (C-C motif) ligand 1 (CCL-1), a well-characterized chemokine secreted by activated T cells, in the nociceptive transmission remains unclear. We found that CCL-1 was upregulated in the spinal dorsal horn after partial sciatic nerve ligation. Therefore, we examined actions of recombinant CCL-1 on behavioural pain score, synaptic transmission, glial cell function and cytokine production in the spinal dorsal horn. Here we show that CCL-1 is one of the key mediators involved in the development of neuropathic pain. Expression of CCL-1 mRNA was mainly detected in the ipsilateral dorsal root ganglion, and the expression of specific CCL-1 receptor CCR-8 was upregulated in the superficial dorsal horn. Increased expression of CCR-8 was observed not only in neurons but also in microglia and astrocytes in the ipsilateral side. Recombinant CCL-1 injected intrathecally (i.t.) to naive mice induced allodynia, which was prevented by the supplemental addition of N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801. Patch-clamp recordings from spinal cord slices revealed that application of CCL-1 transiently enhanced excitatory synaptic transmission in the substantia gelatinosa (lamina II). In the long term, i.t. injection of CCL-1 induced phosphorylation of NMDA receptor subunit, NR1 and NR2B, in the spinal cord. Injection of CCL-1 also upregulated mRNA level of glial cell markers and proinflammatory cytokines (IL-1ß, TNF-α and IL-6). The tactile allodynia induced by nerve ligation was attenuated by prophylactic and chronic administration of neutralizing antibody against CCL-1 and by knocking down of CCR-8. Our results indicate that CCL-1 is one of the key molecules in pathogenesis, and CCL-1/CCR-8 signaling system can be a potential target for drug development in the treatment for neuropathic pain.