[A Case of Laparoscopic Hartmann's Procedure for Rectal Cancer with Inferior Mesenteric Vein-Left Ovarian Vein Shunt].

A 77-year-old woman presented with a chief complaint of bloody stools. Detailed examination revealed a semi-circumferential type 2 tumor in the lower rectum, and a diagnosis of Group 5, tub1-2, cT3N2aM0, cStage Ⅲb rectal cancer was made. Preoperative abdominal CT scans revealed a shunt in the inferior mesenteric vein and left ovarian vein. Laparoscopic Hartmann's procedure was performed, and when the sigmoid mesentery was moved from the inner side, a shunt flowing from the left ovarian vein to the inferior mesenteric vein in the sigmoid mesentery was found, which was then dissected. The operating time was 253 min, and blood loss was approximately 140 g. There was no postoperative liver dysfunction, and the patient was transferred to another hospital on postoperative day 36. Causes of portal-systemic shunts are portal hypertension occurring due to liver cirrhosis or congenital causes and organ adhesion from abdominal surgery. In this case, there was no liver cirrhosis, and the blockage of the left renal vein perfusion by the superior mesenteric artery may have resulted in congestion and varicose of the left ovarian vein. Furthermore, the shunt with the inferior mesenteric vein may have been formed due to the adhesion of the left ovarian vein after ovariectomy. If preoperative tests reveal varices, a surgical treatment is recommended while keeping in mind the possibility of shunt formation as in this case.

[A Case of Pancreatotomy of the Pancreatic Tail with a Preserved Residual Stomach Using ICG Fluorescence Method for Pancreatic Cancer after Gastrectomy of the Pyloric Side].

A 53-year-old male had a history of gastrectomy of the pyloric side for gastric cancer and Billroth Ⅰ reconstruction done 20 years ago. The patient visited the gastrointestinal internal medical department of our hospital with abdominal pain as the chief complaint. Pancreatic cancer was diagnosed with the help of an abdominal CT, and he was then referred to our department. The preoperative disease stage was cT3, N0, M0, Stage ⅡA. As it was over 20 years since the previous surgery and the preoperative CT showed cardiac branches of the left inferior phrenic artery, we inferred that the residual stomach can be preserved. The blood flow was confirmed by the intraoperative ICG fluorescence method, and we then performed pancreatotomy of the pancreatic tail, preserving the stomach and a splenectomy. The pathologic findings were invasive ductal carcinoma, pT3, N1a, M0, Stage ⅡB, and R0. S-1 was administered orally as postoperative adjunctive chemotherapy. The postoperative course has been favorable without recurrence for 2 years. In case a pancreatotomy of the pancreatic tail is performed for cancer of the pancreatic body after gastrectomy of the pyloric side, it was considered that the intraoperative ICG fluorescence method was useful to confirm the blood flow of the residual stomach.

[Metachronous Gastric Intramural Metastasis following Esophageal Cancer ESD-A Case Study].

We report a case of metachronous gastric intramural metastasis following esophageal cancer endoscopic submucosal dissection( ESD). The patient was an 80s man who was referred to the department of gastroenterology of our hospital for earlystage esophageal cancer by a local physician. ESD was performed for a lesion(Lt, 0-Ⅱa+Ⅱc, cT1N0M0, StageⅠ)located 35- 38 cm from the incisors. Pathologic diagnosis revealed that the lesion was a 2.5×2.0 cm-sized, pSM2, 506 mm, well-differentiated squamous cell carcinoma, ly+, v-, pHM0, pVM0. The patient was indicated for additional treatment, but because the patient requested not to undergo operative treatment, radiation therapy, or chemotherapy, strict follow-upwas performed. Upper endoscopy performed 1 year after ESD revealed the presence of a submucosal tumor(diameter of 5 cm)accompanied by ulceration in the gastric cardia, and biopsy findings led to the diagnosis of squamous cell carcinoma. The patient was referred to our department for operative treatment, and considering the possibility of primary squamous cell carcinoma of the stomach, we performed total gastrectomy(D2 dissection, Roux-en-Y)and cholecystectomy. The pathologic diagnosis was well-differentiated squamous cell carcinoma, ly1, v0, SE, N1. Because esophageal cancer and the tissue type were consistent and the primary locus of the tumor was the submucosal layer, the patient was diagnosed with esophageal cancer with gastric intramural metastasis. We report a rare case of metachronous gastric intramural metastasis of esophageal cancer along with a review of the literature.

[Papillary Carcinoma Arising in Thyroglossal Duct Cyst in the Right Lateral Neck].

A 47-year-old woman was admitted to our institution with the chief complaint of a right cervical mass. Imaging examination findings showed a cystic mass of 25mm with a nodular lesion in the right cervical region. Therefore, we performed extirpation of the right cervical cystic mass to allow diagnosis of the lesion. The histopathological findings showed a partial thyroid tissue on the cyst wall covered with glandular epithelium or metaplastic squamous epithelium, and tumor cells proliferating in the papillary form. Considering the histopathological evidence of the characteristic epithelium of the thyroglossal duct cyst, the potential carcinogenesis from the remnant thyroid tissues, and the absence of primary tumor in the thyroid gland, the patient was diagnosed with thyroid papillary carcinoma arising from the thyroglossal duct cyst in the right lateral cervical region. We found recurrence of the right cervical lymph node at 1 year and 5 months after the initial operation. Thus, we performed dissection of the right cervical lymph nodes. Two years and 10 months after the operation, neither recurrence nor metastasis have been observed. It was suggested that, thyroid papillary carcinoma arising from the thyroglossal duct cyst should be taken into consideration when a lateral cervical mass lesion is found.

Salmon cartilage proteoglycan suppresses mouse experimental colitis through induction of Foxp3+ regulatory T cells.

Proteoglycans (PGs) are complex glycohydrates which are widely distributed in extracellular matrix (ECM). PGs are involved in the construction of ECM, cell proliferation and differentiation. ECM components are involved in transduction of proinflammatory responses, but it is still unknown whether PGs are involved in inflammatory response. In this study, we investigated the effect of PG extracted from salmon cartilage on the progression of experimental colitis-induced in severe combined immunodeficiency mice by cell transfer from interleukin-10 (IL-10)-/- mice. IL-10-/- cell-transferred mice showed weight loss, colon shortening and histological appearance of mild colitis. Daily oral administration of PG attenuated the clinical progression of colitis in a dose-dependent manner. Colitis-induced mice showed the elevated expression of IFN-γ, IL-12, TNF-α, IL-21, IL-23p19, IL-6, IL-17A and retinoic acid-related orphan receptor γt (RORγt) in lamina propria mononuclear cells (LPMCs) and oral administration of PG suppressed the expression of these factors. Conversely, expression of Foxp3 that induces CD4+CD25+ regulatory T cells in LPMCs was enhanced by PG administration. These findings suggested that salmon PG attenuated the progression of colitis due to suppression of inflammatory response by enhancement of regulatory T cell induction.

Characterization of heparan sulfate on hepatocytes in regenerating rat liver.

BACKGROUND/PURPOSE: Liver regeneration occurs through interactions between the receptors on hepatocytes, including proteoglycans (PGs) and glycosaminoglycans (GAGs), and various growth factors. We investigated serial changes in GAGs, particularly heparan sulfate (HS), in proliferating hepatocytes. METHODS: We performed 70% hepatectomy in male Wistar rats, and we then isolated hepatocytes by a collagenase perfusion method after each surgery. DNA synthesis was evaluated by measuring proliferating cell nuclear antigen (PCNA). After we had treated the hepatocytes by delipidation and digestion with actinase E, endo-beta-xylosidase, and alpha-amylase, we quantified GAGs by a carbazole-sulfuric acid method. GAGs were analyzed by ion-exchange chromatography, and changes in molecular weight of the HS component were investigated by size-fractionation HPLC. RESULTS: Hepatocyte mitosis peaked at 24 h after the amount of GAGs was increased at 24 and 72 h after surgery. The amount of HS was slightly increased at 3 to 12 h after surgery, and then peaked at 24 h. The molecular weight of the HS declined by 12 h, but had recovered to the preoperative level by 24 h. CONCLUSIONS: These results suggested that this HS molecule, which contained about ten disaccharide units during proliferation, may be an initiator of hepatocyte proliferation.

Effects of proteoglycan on dextran sulfate sodium-induced experimental colitis in rats.

Proteoglycans (PG) are macromolecules composed of glycosaminoglycan chains covalently attached to a protein core. In this study, we examined the effects of PG on dextran sulfate sodium (DSS)-induced experimental colitis in rats. First, to examine whether PG may ameliorate acute established DSS colitis, PG was administered orally for 5 days to the model animals. We evaluated the effects of PG on the basis of clinical symptoms, hematological analysis, macroscopic observation, and microscopic examination. We then examined whether PG administered orally to rats was detectable in their colonic lumen. After administration of PG, the colonic contents were collected, and the molecular weight of PG in the sample was analyzed by gel filtration high-performance liquid chromatography. Furthermore, we examined whether orally administered PG affected the concentrations of short-chain fatty acids (SCFAs) in the colonic feces. Orally administered PG ameliorated the clinical symptoms of bloody stools and diarrhea, and attenuated the increase in the white blood cell count in rats with established DSS colitis. Histologically, orally administered PG reduced the degree of mucosal erosion and inflammatory cell infiltration into the erosive area induced by DSS. Orally administered PG was detected in rat colon, although its molecular weight was slightly decreased. Orally administered PG significantly increased the concentration of total SCFAs and n-butyrate in rat colonic feces. This is the first study to indicate that exogenous PG ameliorates experimental colitis, suggesting the potential usefulness of PG for clinical treatment of colitis.

Decreased expression of claudin-1 correlates with recurrence status in breast cancer.

Claudins (CLDNs) constitute the major transmembrane proteins of tight junctions. It may be hypothesized that changes in or loss of expression of tight junctional proteins such as CLDNs can lead to cellular disorientation and detachment, which is commonly seen in neoplasia. Recent studies have suggested that claudin-1 (CLDN1) plays an important role in invasion and metastasis and claudin-4 (CLDN4) has a particular role in mammary glandular cell differentiation and carcinogenesis. In this study, we examined 83 breast cancer cases and demonstrated immunohistochemical expression patterns of CLDN1/CLDN4 in recurrent and non-recurrent groups. We found significant results between the recurrent and non-recurrent group for expression of CLDN1/CLDN4. The recurrent group (26 cases) showed decreased expression patterns of CLDN1 (p<0.001), compared to the non-recurrent group (57 cases). Decreased expression of CLDN1 (p<0.0001) correlated with short disease-free interval. The lymph node metastasis-positive group showed decreased expression patterns of CLDN1 (p=0.001). However, there was no significance between the recurrent group and non-recurrent group in CLDN4 expression. There was no significance between histological factors and CLDN4 expression. The results indicated that CLDN1 expression correlated with the recurrence status and malignant potential of breast cancer.

Suppression of matrix metalloproteinase-9 by 4-methylumbelliferone.

OHK cells, a human lymphoma cell line, are known to produce large amounts of hyaluronan. We investigated the effect of 4-methylumbelliferone, an inhibitor of hyaluronan synthesis, on the activity of matrix metalloproteinases in OHK cells. Matrix metalloproteinase-9 was detected on gelatin zymography as the main metalloproteinase excreted into the medium of cultured OHK cells, and 4-methylumbelliferone added to the medium decreased the activity of the enzyme in a dose-dependent manner. Addition of Streptomyces hyaluronidase to the medium during cultivation did not decrease the enzyme activity. Reverse transcription-polymerase chain reaction revealed that 4-methylumbelliferone markedly decreased the level of mRNA for matrix metalloproteinase-9 in cultured OHK cells. A similar decrease of the activity of matrix metalloproteinase-9 by 4-methylumbelliferone was also observed in cultured human breast and colon carcinoma cells. These results suggest that 4-methylumbelliferone suppresses the expression of matrix metalloproteinase-9 in cultured cancer cells.

Plasma exchange-based plasma recycling dialysis system as a potential platform for artificial liver support.

We developed a plasma recycling dialysis (PRD) system based on plasma exchange (PE). In this system, rapid reduction of toxic substances and restitution of deficient essential substances are performed by PE, and subsequent blood purification is performed by dialysis between separated plasma recycled over a purification device and the patient's blood across the membrane of the plasma separator. This study was performed to demonstrate the safety and efficacy of this system. Hyperbilirubinemia was induced by ligating the bile duct in pigs, and 7 days later, only PE for 2 h (group PE) or PE for 2 h followed by PRD for 6 h (group PE + PRD) was performed. The separated plasma was recycled over anion-exchange resin through the extra fiber space of the plasma separator. The safety and efficacy of this system were evaluated based on the values of hemodynamic and laboratory parameters. Transfer from PE to PRD was completed in a few minutes. The hemodynamic status and blood cells counts were stable and hemolysis was not observed during the procedure. In the PE + PRD group, the concentrations of total bile acids continuously decreased (pretreatment, 155.5 +/- 40.6 microM; 2 h [end of PE], 76.1 +/- 14.4 microM; 8 h [end of PRD], 25.8 +/- 9.1 microM) and the value was significantly lower than in the PE group after 6 h. The total bilirubin also continuously decreased during PRD (pretreatment, 55.3 +/- 11.5 microM; 2 h [end of PE], 33.8 +/- 8.4 microM; 8 h [end of PRD], 18.6 +/- 7.7 microM) and was significantly lower than in the PE group after 4 h. No significant change was observed in other laboratory values. This PE-based PRD system allowed a swift transfer from PE to sorbent-based blood purification. The safety of this system was demonstrated and the removal of toxic substances was significant. This study confirmed the clinical utility of this system as a platform for artificial liver support.

Study of hyaluronan synthase inhibitor, 4-methylumbelliferone derivatives on human pancreatic cancer cell (KP1-NL).

The structure of 4-methylumbelliferone (MU) consists of coumarin with 4-methyl group and 7-hydroxy group. MU inhibits HA synthesis and pericellular HA matrix formation. In this study, we used 10 MU derivatives which have hydroxy groups and methyl groups at various positions of coumarin to investigate a more effective HA inhibitor than MU. First, human pancreatic cancer cell (KP1-NL) growth assay was analyzed by Alamar Blue to determine the non-toxic concentration of MU derivatives, and the inhibitory effect on HA synthesis in the cell cultures was analyzed by HA measuring kit. Next, cell surfaces of cancer cells were analyzed by particle-exclusion assay. In conclusion, both hydroxy and methyl groups are necessary for HA inhibition by MU, and two hydroxy groups inhibited HA synthesis more strongly than MU.

4-methylumbelliferone, a hyaluronan synthase suppressor, enhances the anticancer activity of gemcitabine in human pancreatic cancer cells.

Hyaluronan (HA) is a ubiquitous, major component of the pericellular matrix and is necessary for various physiological processes. It plays a very important role in biological barriers. We previously reported that 4-methylumbelliferone (MU) inhibits HA synthesis and pericellular HA matrix formation in cultured human skin fibroblasts, Streptococcus equi FM100, and B16F10 melanoma cells. We hypothesized that MU-mediated inhibition of HA synthesis and pericellular HA matrix formation would increase the efficacy of anticancer drugs. We have already demonstrated in vitro, using a sandwich binding protein assay and a particle exclusion assay, that MU inhibits HA synthesis and formation of the pericellular HA matrix, respectively, in human KP1-NL pancreatic cancer cells. AlamarBlue assay revealed that the anticancer effect of gemcitabine in KP1-NL cells was increased by pretreatment with MU. In vivo simultaneous administration of MU and gemcitabine to tumor-bearing mice with severe combined immunodeficiency disease (SCID) decreased the size of the primary and metastatic tumors more than did gemcitabine alone. These data strongly suggest that a combination of MU and gemcitabine is effective against human pancreatic cancer cells. MU may have potential as a chemosensitizer and may provide us with a new anticancer strategy.

A hyaluronan synthase suppressor, 4-methylumbelliferone, inhibits liver metastasis of melanoma cells.

4-Methylumbelliferone (MU) inhibits the cell surface hyaluronan (HA) formation, and that such inhibition results in suppression of adhesion and locomotion of cultured melanoma cells. Here, we examine the effect of MU on melanoma cell metastasis in vivo. MU-treated melanoma cells showed both decreased cell surface HA formation and suppression of liver metastasis after injection into the mice. Oral administration of MU to mice decreased tissue HA content. These HA knock-down mice displayed suppressed liver metastasis. Thus, both cell surface HA of melanoma cells and recipient liver HA can promote liver metastasis, indicating that MU has potential as an anti-metastatic agent.

Effect of a hyaluronan synthase suppressor, 4-methylumbelliferone, on B16F-10 melanoma cell adhesion and locomotion.

Hyaluronan (HA) is a ubiquitous, major component of the extracellular matrix. It is involved in cell adhesion and locomotion, and hence in tumor metastasis. We have previously reported that 4-methylumbelliferone (MU) inhibits HA synthesis and may be a useful tool for examining the functions of HA. We here demonstrate that the formation of cell surface HA by melanoma cells and its release into the culture medium are inhibited by MU. Adhesion and locomotion assays revealed that the adhesion and locomotion of melanoma cells were dose-dependently inhibited by MU. Conversely, treatment with exogenous HA enhanced both adhesion and locomotion. Thus, preventing the formation of cell surface HA reduced both the adhesion and locomotion of melanoma cells, suggesting that MU may act as an inhibitor of tumor metastasis.

Neoplastic changes in saccharide sequence of dermatan sulfate chains derived from human colon cancer.

Decorin, a small proteoglycan containing a dermatan sulfate (DS) chain, is expressed abnormally in human colon cancer stroma. The aim of this study was to determine neoplastic changes in DS chains from human colon cancer and normal colonic mucosa. Proteoglycans were extracted from human colon cancer and normal colonic mucosa and successively digested with enzymes. The glycosaminoglycan obtained was fluoro-labeled with 2-aminopyridine at reducing terminals and fractionated by HPLC. Fluoro-labeled DS chains were collected and digested with bovine testicular hyaluronidase, followed by HPLC. The repeating disaccharide connected to the linkage region [glucuronosyl-galactosyl-galactosyl-xylosyl(2-aminopyridine)] of pyridylaminated DS chains from both types of tissue was glucuronosyl-N-acetylgalactosamine. The other glucuronic acid of the pyridylaminated DS chain was located 12 saccharides from the reducing terminal in colon cancer, and 18 saccharides from the reducing terminal in normal colon. The saccharide sequence of DS chains from human colon cancer is altered from that in normal colon.