[A case of liver Mycobacterium avium complex syndrome, diagnosed by liver biopsy].

A man in his 70s was referred to our hospital for evaluation of low-grade fever, weight loss, and liver dysfunction. Serological tests for viral hepatitis or autoimmune diseases were negative. No significant findings were observed on whole-body computed tomography (CT). Histopathologic examination of a liver biopsy sample revealed a non-caseating granuloma with acid-fast bacillus using the Ziehl-Neelsen stain. Serum Mycobacterium avium complex (MAC) antibody was positive. We started treatment for pulmonary MAC disease. His clinical condition and liver function improved within two months. He was diagnosed with liver MAC disease.

Increased Insulin Secretion from Insulin-Secreting Cells by Construction of Mixed Multicellular Spheroids.

PURPOSE: We previously have shown that multicellular spheroids containing insulin-secreting cells are an effective therapy for diabetic mice. Here we attempted to increase insulin secretion by incorporating other cell types into spheroids. MATERIALS AND METHODS: Multicellular spheroids of mouse MIN6 pancreatic ß cells were formed in microwells alone and with aortic vascular endothelial MAEC cells or embryo fibroblast NIH3T3 cells. mRNA expression of insulin genes and insulin secretion of MIN6 cells in each spheroid were measured by real-time PCR and an insulin ELIZA kit. Moreover, collagen IV expression in each spheroid was analyzed by western blot. RESULTS: In all cases, uniformly sized (about 300 µm) multicellular spheroids were obtained. MAEC or NIH3T3 cell incorporation into MIN6 spheroids significantly increased mRNA expression of insulin genes and insulin secretion. In addition, collagen IV expression, which was reported to enhance insulin secretion from pancreatic ß cells, also increased in their spheroids. CONCLUSIONS: The formation of mixed multicellular spheroids containing collagen IV-expressing cells can improve the insulin secretion from insulin-secreting MIN6 cells, and mixed multicellular spheroids can be a potent therapeutic option for patients with type I diabetes mellitus.

[A recurrent gastric cancer with peritoneal metastasis 15 years after distal gastrectomy successfully treated with S-1 and weekly intravenous and intraperitoneal injections of Paclitaxel].

Fifteen years after receiving a distal gastrectomy for advanced gastric cancer, a 70-year-old woman was admitted to our hospital because of abdominal fullness due to ascites. Although cytological examination showed adenocarcinoma cells in the fluid, no examination revealed the primary lesion. Peritoneal metastasis was detected via immunohistochemistry using the cell block technique. After chemotherapy failure (S-1 plus CDDP, weekly PTX, and S-1 plus DOC), the patient received S-1 and weekly intravenous and intraperitoneal injections of PTX. The ascites decreased, and she has been doing well. Our experience with this case suggests that S-1 and weekly intravenous and intraperitoneal injections of PTX is a promising means of treating gastric cancer with peritoneal metastasis.

[A case of superior mesenteric artery occlusion with ischemic enteritis after transcatheter therapy].

An 84-year-old female was admitted with sudden-onset upper abdominal pain. Contrast-enhanced computed tomography (CECT) revealed complete occlusion of the superior mesenteric artery (SMA). After transcatheter infusion of urokinase, embolic occlusion resolved. However, the pain recurred when she started eating. CECT revealed a lesion with thickening of the intestinal wall; therefore, laparoscopy-assisted surgery was undertaken. Histological examination yielded a definitive diagnosis of ischemic enteritis caused by SMA occlusion. Rapid diagnosis and treatment are important in SMA occlusion, and careful observation of the clinical course is recommended after transcatheter therapy.

Transplantation of insulin-secreting multicellular spheroids for the treatment of type 1 diabetes in mice.

The efficacy of cell-based therapy depends on the function and survival of transplanted cells, which have been suggested to be enhanced by spheroid formation. However, few attempts at spheroid generation from insulin-secreting cells, which may be used to treat type 1 diabetes, have been reported. We therefore developed spheroids from the mouse insulinoma cell line NIT-1 by using polydimethylsiloxane (PDMS)-based microwells with a coating of poly(N-isopropylacrylamide) (PNIPAAm). The prepared NIT-1 spheroids or dissociated NIT-1 cells were transplanted into the subrenal capsule in streptozotocin-induced diabetic mice. NIT-1 spheroids prepared using the PNIPAAm-coated PDMS-based microwells had a uniformly sized spherical structure with a diameter of 200-300µm. The PNIPAAm coating increased cell survival in the spheroids and the recovery of the spheroids from the microwells. In diabetic mice, the transplanted NIT-1 spheroids reduced blood glucose levels to normal values faster than dissociated NIT-1 cells did. Additionally, survival was higher among NIT-1 cells in spheroids than among dissociated NIT-1 cells 24h after transplantation. These results indicate that insulin-secreting NIT-1 spheroids prepared using PNIPAAm-coated PDMS-based microwells are more effective for the treatment of type 1 diabetes than dissociated cells in suspension.

[Pancreatic pseudocyst with fistula to the common bile duct and the duodenum].

A 35-year-old man was hospitalized for severe acute pancreatitis. On the 24th hospital day, CT scan showed a pancreatic pseudocyst in the head of the pancreas. Conservative medical treatment for 1 month was not effective, and CT scan revealed a fistulous communication of the pseudocyst to the common bile duct and duodenum. After the formation of a fistulous communication, we detected common bile duct stones composed of fatty acid calcium and we removed them endoscopically. The pseudocyst gradually decreased in size and disappeared 4 months later. Follow-up CT scan showed no sign of recurrence.

Poly(N-isopropylacrylamide)-coated microwell arrays for construction and recovery of multicellular spheroids.

Microwell arrays that have many micro-sized cavities on the device have been employed to form multicellular spheroids. However, methods to efficiently harvest the constructed spheroids from the microwell arrays have not been thoroughly investigated. We evaluated the effects of poly(N-isopropylacrylamide) (PNIPAAm) for constructing and harvesting spheroids from microwell arrays. Microwell arrays were coated with ethanol containing 1%, 2.5%, 5%, or 10% PNIPAAm by a solvent-casting method and then dried. Spheroids formed using the coated microwell arrays were harvested. Highly efficient and rapid recovery of NIH3T3 mouse fibroblast spheroids were achieved for the 5% and 10% coated wells (93.2% ± 1.6% and 93.6% ± 1.1% at 60 s, respectively), whereas recovery was not efficient for 0%, 1%, and 2.5% coated wells (0.2% ± 0.2%, 1.1% ± 0.6%, and 7.8% ± 4.0% at 60 s, respectively). Because PNIPAAm is a thermoresponsive polymer that exhibits a lower critical solution temperature (LCST) of 32°C, we examined the effects of temperature on the recovery rate. The recovery rates at 4°C (below LCST) were equivalent to or higher than those at 37°C (above LCST) for all four cell types examined. Functional assessment suggests that the PNIPAAm microwell arrays are not toxic to the formed spheroids. The PNIPAAm microwell array developed in the present study will be useful for constructing and harvesting spheroids.

[A case of small pancreatic cancer with intraductal progress].

A 60-year-old man was admitted to our department for further evaluation of main pancreatic duct dilatation detected on ultrasonography. Endoscopic retrograde cholangiopancreatography showed stenosis of the main pancreatic duct at the junction of the pancreatic head and body. Brush cytology revealed pancreatic ductal carcinoma. Histological examination of the resected pancreas showed a 15-mm in length intraductal growth of carcinoma in situ in the main pancreatic duct, 10mm of which showed microinvasion. There was also atypical hyperplasia at a branch pancreatic duct near the lesion. It was suspected to be an initial stage of pancreatic ductal carcinoma. Intraductal progression type and non-progression type are often suspected in a case showing progression of carcinoma in situ to pancreatic ductal carcinoma, and this case was thought to be intraductal progression.

The cytological characteristics of small cell change of dysplasia in small hepatic nodules.

Small cell change of dysplasia (SCD) is characterized as an initial step in hepatocarcinogenesis. Histopathological diagnosis is an important diagnostic procedure for nodular lesions in the liver. However, the biopsied specimen is so small that it is sometimes difficult to differentiate between regenerative nodules, dysplastic nodules, and hepatocellular carcinoma even histologically. To examine the usefulness of cytology in the differential diagnosis of hepatic nodular lesions, the cellular characteristics of SCD were evaluated using Papanicolaou staining and a micrometer. Sixty-four histologically diagnosed small nodular lesions in the liver were analyzed retrospectively. All cases were histologically classified according to the Terminology of Nodular Hepatocellular Lesions by the International Working party: hepatocellular carcinoma (HCC) (n=17); low-grade dysplastic nodule (LGDN) (n=26); high-grade dysplastic nodule (HGDN) (n=6); large regenerative nodule: (n=15). SCD was noted in all of the histological categories, and the proportion of SCD tended to be higher in W-HCC than in dysplastic nodules. Although the cellular size was the smallest in HGDN, the nuclear size was the largest in well-differentiated HCC (W-HCC). The nuclear/cytoplasmic ratio was higher in HGDN and W-HCC than in other nodular lesions. Hyperchromasia in W-HCC was obviously stronger than that in other nodules. SCD was frequently found in HGDN and W-HCC. The present study showed that detailed cytological findings of SCD are useful for differentiating HGDN from LGDN, and HGDN from W-HCC.

Complete response of a pancreatic adenosquamous carcinoma to chemoradiotherapy.

A 51-year-old woman with an unresectable pancreatic tumor that was histologically diagnosed as an adenosquamous carcinoma underwent chemoradiotherapy with 5-fluourouracil (FU) and low-dose cisplatin (low-dose FP). Because we recognized a partial response to the chemoradiotherapy, we subsequently administered combined chemotherapy with S-1 and cisplatin. After one course of this combined chemotherapy, the tumor was further reduced in size and became difficult to discern on abdominal computed tomography (CT). We have continued to administer the S-1 and cisplatin combined chemotherapy, and the patient is still alive. After 20 months of treatment, the tumor has not recurred (as assessed by abdominal CT). Additionally, we have not seen elevation of tumor markers. This report presents the successful use of chemoradiotherapy with low-dose FP and additional combined chemotherapy with S-1 and cisplatin for unresectable pancreatic adenosquamous carcinoma.