Quantitative study on appearance of microvessels in spectral endoscopic imaging.

Increase in abnormal microvessels in the superficial mucosa is often relevant to diagnostic findings of neoplasia in digestive endoscopy; hence, observation of superficial vasculature is crucial for cancer diagnosis. To enhance the appearance of such vessels, several spectral endoscopic imaging techniques have been developed, such as narrow-band imaging and blue laser imaging. Both techniques exploit narrow-band blue light for the enhancement. The emergence of such spectral imaging techniques has increased the importance of understanding the relation of the light wavelength to the appearance of superficial vasculature, and thus a new method is desired for quantitative analysis of vessel visibility in relation to the actual structure in the tissue. Here, we developed microvessel-simulating phantoms that allowed quantitative evaluation of the appearance of 15-µm-thick vessels. We investigated the relation between the vascular contrast and light wavelength by the phantom measurements and also verified it in experiments with swine, where the endoscopically observed vascular contrast was investigated together with its real vascular depth and diameter obtained by microscopic observation of fluorescence-labeled vessels. Our study indicates that changing the spectral property even in the wavelength range of blue light may allow selective enhancement of the vascular depth for clinical use.

[Complete response in a case of advanced unresectable esophageal cancer treated by chemoradiation therapy and S-1+CDDP chemotherapy].

We report a 37-year-old woman who complained of chest discomfort as of August 2004, and was found to have advanced esophageal cancer in the upper thoracic area in December 2004.S he was diagnosed as Stage IVa (T4N1M0) because chest computed tomography (CT) indicated trachea invasion and lymph node metastasis. We diagnosed it to be a case of unresectable esophageal cancer, and she underwent chemoradiation therapy. CT showed regression of the main tumor and metastatic lymph nodes when the CRT course was completed. The main tumor disappeared macroscopically. We again considered an operation, but the CRT was so effective that the patient wished to continue CRT and underwent three courses. Endoscopy showed disappearance of the main tumor and Lugol's solution. Following this, 10 courses of the treatment with CDDP alone (CDDP 10 mg/weekly) were continued until the appearance of renal dysfunction. S-1 (100 mg/body/day)was started in September 2005. The treatment is currently ongoing, and no recurrence or metastases had occurred as of March 2009.

[A case of recurrent pancreatic cancer with lung metastasis responding to S-1 combined gemcitabine chemotherapy].

The patient was a 54-year-old male. In July 2004, he underwent resection of the pancreatic body tail region to treat pancreatic body tail cancer. On histopathological examination, the stump of the extirpated specimen was positive for tumor cells. After surgery, 10 courses of therapy with gemcitabine hydrochloride(GEM, 1, 000 mg/m(2), 3-week administration followed by 1-week discontinuation)were performed, and follow-up was continued. In February 2006, local relapse was detected. Chemotherapy with GEM was administered for 1 year and 9 months. However, in November 2007, an increase in the recurrent lesion size and right lung metastasis were noted. The regimen was switched to combination therapy with S-1 and GEM(S-1 60 mg/m(2) day, continuous administration on days 1 to 14 and 2-week discontinuation; and GEM 1, 000 mg/ m(2), administered on days 8 and 15). After the end of the 11th course, PET-CT revealed the disappearance of FDG accumulation in the recurrent and metastatic lesion sites. During the treatment period, there were no grade 3 or higher adverse reactions. The patient is being treated at the outpatient clinic (as of January 2009).

[A case of Stage IV non-operated advanced breast cancer with local recurrence thirteen years after initial therapy].

A 73-year-old woman underwent chemotherapy, radiotherapy, and hormonal therapies for right advanced breast cancer (T4, N3, M1, Stage IV) in 1993. She obtained a complete response in pathological evaluation (pCR) in 1998. For adjuvant therapies, she had been treated with hormonal therapies for five years until 2003. After the interruption of hormonal therapies, the serum levels of tumor markers had been elevated, and she had a right axillary local recurrence and a right ovarian metastasis detected by a FDG-PET/CT in February 2006. The right axillary local recurrence lesion was then resected, and she has since been treated with hormonal therapy of aromatase inhibitor (AI). The serum levels of tumor markers have been remarkably reduced, and FDG-PET/CT has showed the disappearance of the right axillary local recurrence, and the decrease of FDG accumulation of the right ovarian metastases in February 2007. We present a case of non-operated advanced breast cancer with local recurrence surviving successfully long term when treated with AI.

[A case of advanced pancreatic cancer successfully treated by combined chemotherapy of S-1 and gemcitabine as second-line chemotherapy].

The patient was a 63-year-old man who suffered from advanced pancreatic cancer (T 4 N 3 M 0, Stage IVb). Palliative operation was performed for obstructive jaundice. He was treated with chemotherapy of gemcitabine (GEM) alone as first-line, and combined chemotherapy of GEM and S-1 as second-line. Both therapies were effective for this patient. Tumor marker (CA 19-9) decreased after chemotherapies (first-line: 5,692 U/mL to 70 U/mL, second-line: 4,877 U/mL to 562 U/mL). No toxic events were observed due to these therapies, so he was treated as an outpatient for about 2 years. It was considered that he had a good quality of life.

[Weekly paclitaxel therapy effective for gastric cancer with obstructive jaundice due to peritoneal dissemination--a case report].

The patient was a 61-year-old man who suffered from advanced gastric cancer, and a distal gastrectomy was performed (T3N2P1CY1, Stage IV). He was treated with chemotherapy of TS-1 alone (100 mg/day, days 1-28 with two weeks rest). Six months later he complained of lumbago and appetite loss, then was admitted to the hospital with obstructive jaundice. Total bilirubin (T-Bil) was increased to 11.3 mg/dl. CT scan examination revealed peritoneal dissemination with much ascites and dilatation of intrahepatic bile ducts. Endoscopic drainage was tried, but was discontinued due to stenosis of gastroduodenal anastomosis. Ultimately, T-Bil was elevated to 25.2 mg/dl, and he could not sleep comfortably because of a severe itch and an irritating feeling. Weekly paclitaxel therapy was started (70 mg/m(2), day 1, 8, 15, once a week for 3 weeks followed by a week rest as one cycle). One month after the first infusion therapy, the obstructive jaundice was notably improved and the ascites disappeared completely, so he was discharged. For about one year, he was treated with this chemotherapy as an outpatient. The toxic events were anemia (grade 3) and alopecia (grade 1).

Protective effects of nafamostat mesilate on liver injury induced by lipopolysaccharide in rats: possible involvement of CD14 and TLR-4 downregulation on Kupffer cells.

Nafamostat mesilate (NM) is a synthetic protease inhibitor with various biological effects. To determine its effect on liver injury related to sepsis, we investigated the effects of NM on lipopolysaccharide (LPS)-induced liver injury. Wistar rats were allocated into two groups; the NM group underwent intraperitoneal NM administration 30 min before LPS administration, and the control group underwent PBS administration. Serum AST and ALT levels were significantly decreased in NM-treated rats. Reduced levels of TNF-alpha, IL-1beta, and IFN-gamma were observed after LPS administration in NM-treated rats. No significant differences were observed in IL-6 levels between the NM and the control group. In contrast, HGF levels were significantly increased only in control rats. NM treatment decreased protein and mRNA levels of TLR-4 and CD14. Our data suggest that NM treatment has protective effects against LPS-induced hepatotoxicity through downregulation of TLR4 and CD14 in liver, which decreased TNF-alpha, IL-1beta, and IFN-gammaproduction in liver.

Obstructive jaundice increases sensitivity to lipopolysaccharide via TLR4 upregulation: possible involvement in gut-derived hepatocyte growth factor-protection of hepatocytes.

BACKGROUND: Patients with obstructive jaundice are prone to sepsis after biliary tract surgery. The present study was designed to determine the effect of biliary obstruction on cytokine responses to lipopolysaccharide (LPS). METHODS: Wister rats were allocated into two groups; the BDL group underwent bile duct ligation, followed 2 weeks later by administration of LPS into the duodenum. The control group underwent sham operation, and similarly received enteral LPS. Specimens were collected serially, and applied for the assays. RESULTS: Serum aspartate aminotransferase and alanine aminotransferase levels were significantly increased in BDL rats. High tumor necrosis factor alpha (TNF-alpha) and interleukin (IL)-6 levels in peripheral blood were observed 2 h after LPS administration in BDL rats. In contrast, no increases in both cytokines were noted in peripheral and portal blood in control rats. Baseline HGF levels in portal and peripheral blood in BDL rats were significantly higher than in control rats. LPS significantly increased hepatocyte growth factor (HGF) levels in portal blood and decreased in peripheral blood in BDL rats, but not in control rats. Immunohistochemical analysis revealed that BDL increased expressions of Toll-like receptor (TLR)4, CD14 and CD68 both in the small intestine and liver. Both TLR4 and CD14 mRNAs were upregulated in the small intestine and liver after LPS administration in BDL rats. CONCLUSION: Obstructive jaundice and LPS stimulation induced TLR4 upregulation both in the liver and small intestine, which led to increased TNF-alpha and IL-6 production in liver and HGF production in the small intestine. The upregulation of TLR4 may lead to pathological and host defense reactions in obstructive jaundice complicated with Gram-negative bacterial infection.

Failure of the gut barrier system enhances liver injury in rats: protection of hepatocytes by gut-derived hepatocyte growth factor.

OBJECTIVE: Clinical and experimental studies suggest that impairment of the mucosal barrier system increases gut-derived endotoxin in the portal blood, which causes liver injury. The aim of this study was to elucidate the mechanism of liver injury caused by gut defence failure. DESIGN: Wistar rats were administered either enteral lipopolysaccharide (LPS) or LPS via the portal vein. METHODS: Blood samples were collected via the inferior vena cava at necropsy. Serum aspartate transaminase (AST) and alanine transaminase (ALT) were analysed by standard enzymatic procedures and cytokines [tumour necrosis factor-alpha, interleukin (IL)-1beta, interferon-gamma, IL-6 and hepatocyte growth factor (HGF)] were measured by enzyme-linked immunosorbent assay. Livers were removed and snap-frozen in liquid nitrogen. CD14, CD68, Toll-like receptor (TLR) 2, TLR4 and Fas ligand (FasL) were analysed immunohistochemically. Expression of TLR2, TLR4 and CD14 mRNA was determined by reverse transcriptase-polymerase chain reaction. RESULTS: In enterally-treated rats, AST and ALT were not increased and cytokine levels were under the limits of detection in the absence of a rise in HGF. Enteral administration of LPS increased HGF dose-dependently. Injection of LPS in the portal vein resulted in significant increases in AST, ALT, tumour necrosis factor-alpha, IL-1beta, interferon-gamma and IL-6 levels, but no change in HGF levels. Immunohistochemical analysis revealed that intraportal LPS administration increased CD14, TLR4, CD68 and FasL. Reverse transcriptase-polymerase chain reaction analysis demonstrated that TLR4 mRNA expression was upregulated 0.5 h after intraportal LPS administration. CONCLUSION: s Our data suggest that Kupffer cell activation mediated by intraportal LPS via TLR4 is involved in liver injury, possibly through both tumour necrosis factor-alpha/IL-1beta and FasL, and that lack of HGF activity in the impaired gut could not counteract liver injury.

The impact of portal infusion with donor-derived bone marrow cells and intracellular cytokine expression of graft-infiltrating lymphocytes on the graft survival in rat small bowel transplant model.

BACKGROUND: Intraportal administration of alloantigen is reported to reduce antigen-specific immune responses, although the underlying mechanisms for the reduced immunological reactions, especially those of the graft, are poorly understood. We examined intracellular cytokine production by graft-infiltrating lymphocytes (GILs) and peripheral blood lymphocytes (PBLs) to elucidate the underlying mechanisms of beneficial effects on intraportal infusion of donor cells in rat small bowel transplantation (SBT). METHODS: Recipient rats (Lewis) were transplanted with small bowel from ACI rats. Tacrolimus (Tac) was injected daily from days 0 to 4. Bone marrow cells of ACI rats were infused via the portal or tail vein on the day of surgery. On day 5, both GILs and PBLs collected from SBT graft and peripheral blood, respectively, were analyzed for intracellular cytokine production of recipient-derived alphabeta-T cells. The Th1/Th2 balance in each group was designated as the ratio of the percentage of GILs or PBLs staining positive for intracellular IL-4 or IFN-gamma, respectively. The total cell numbers of GILs from SBT graft were also counted. RESULTS: Survival of recipients was markedly prolonged by the combination of Tac and donor-specific bone marrow infusion via the portal vein (DSBMI-PV-Tac) compared with the untreated control, Tac alone, or DSBMI tail vein plus Tac. DSBMI-PV-Tac significantly decreased the total cell numbers of GILs and also induced remarkable Th2-type response in GILs. CONCLUSIONS: Our results indicate that DSBMI-PV-Tac decreased GILs and enhanced Th2-type response in SBT graft, both of which are associated with a significant prolongation of graft survival in SBT.