Background. 　Although the development of a practicable culture method to maintain long-term hepatocyte function is highly desirable, hepatocytes lose their liver-specific functions rapidly on regular collagen-coated dishes. In this study, we evaluated the efficacy of a special cell culture dish coated with the temperature-responsive polymer poly(N-isopropylacrylamide) (PIPAAm), and observed the morphological and functional changes of hepatocytes on changing the amount of polymer. Methods. 　Culture plates with varying amounts of PIPAAm polymer (1.5〜3.5 times that in the commercially available temperatureresponsive polymer-containing dish (UpCell^®, CellSeed, Tokyo, Japan)) were prepared. All dishes were enhanced by adding a layer of rat tail collagen I at a dose of 600 μg/dish for 3 hours. Hepatocytes isolated from Sprague-Dawley rats were cultured in these static culture systems. Morphologic changes and liver-specific functions were evaluated. Results. 　On day 7 of culture, spheroid formation of hepatocytes was observed in the high-polymer group, with the presence of glycogen and albumin in the spheroid. In the high-polymer group, the rate of albumin production was significantly higher than in the low-polymer group until day 14 of culture (P < 0.001). Conclusion.　 The spheroid formation of hepatocytes cultured in the presence of a high level of PIPAAm showed the long-term maintenance of liver-specific functions in vitro.

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AUF1 is one of Adenine-and-Uridine-rich elements binding protein which regulates the mRNA stability of many genes related to growth regulation, cytokines, and cell cycle-regulatory genes. There was no report about the expression of AUF1 in gliomas. Sections of formalin-fixed, paraffin-embedded tissues from 71 gliomas were stained immunohistochemically using a polyclonal antibody against the adenine-and-uridine-rich element RNA-Binding factor 1 (AUF1) oncoprotein. Positive staining, which is known to correlate with gene amplification, was not associated with patients’ sex, age, Karnofsky performance status scores (KPS), tumor size, Bcl-2 expression, or longer overall survival. However, positive staining was negatively correlated with the MIB-1 labeling index, while it was positively correlated with the lower grade group of WHO classification. Expression of the AUF1 oncoprotein appears to be an important additional indicator in human gliomas.

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　We have previously shown that wild type (wt) mice exhibit susceptibility to immunization with human (h) thyrotropin receptor (TSHR), but resistance to mouse (m) TSHR, while TSHR knockout (KO) mice are susceptible to mTSHR, indicating the existence of robust immune tolerance against the mTSHR in wt mice. This tolerance may be mediated by either centrally or peripherally. We here explored the contribution of a peripheral arm of immune tolerance against the mTSHR by using antibodies to deplete regulatory T cells (Tregs), to antagonize co-inhibitory molecules and/or to stimulate co-stimulatory molecules. Antagonistic anti-co-inhibitory molecules, cytotoxic T lymphocyte antigen 4 (CTLA4) and programmed cell death 1 ligand 1 (PDL1), induced only low levels of anti-TSHR antibodies without induction of hyperthyroidism in a mouse Graves’ model. In this experimental setting, antibody levels were significantly higher in THSR^+/- mice than wt mice. However, agonistic anti-co-stimulatory molecules, CD40 and CD137, and Treg-depleting anti-CD25 antibodies showed no effect. All these data suggest that peripheral immune tolerance against the mTSHR may play a minor role, and imply the importance of central tolerance, in immune tolerance against mTSHR in mice. Additional studies on central tolerance to the mTSHR will be necessary for completely delineating the mechanisms for immune tolerance against mTSHR in mice.

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Objectives: Reactive oxygen species (ROS) function as key metabolites that can impair biological processes. The aim of this preliminary study was to assess the perioperative oxidative changes in lung cancer surgery. 　Methods: We measured the levels of blood hydroperoxides, a type of ROS, as an index of oxidative injury to cellular components, as well as the plasma ferric-reducing ability as an index of total antioxidant potential in 32 lung cancer patients. Hydroperoxides were measured by the levels of diacron reactive oxygen metabolites (d-ROMs). The antioxidant potential was determined by the biological antioxidant potential (BAP), which represents the levels of endogenous antioxidant enzymes. 　Results: Lung cancer patients had slightly affected oxidative stress before surgery. The d-ROM and BAP levels after surgery and were significantly decreased than before surgery (p < 0.001) and the levels recovered preoperatively at third postoperative day. The d-ROM level in video-assisted surgery group (n=17) was significantly decreased than those in thoracotomy group (n=15) at third and seventh post-operative day (p < 0.001, and < 0.02). 　Conclusions: Lung cancer patients had already exposed oxidative stress before surgery and surgical intervention also generates large amounts of ROS. Video-assisted thoracic surgery can reduce the ROS compared to the standard thoractomy.

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A 73-year-old woman was found to have diabetes mellitus along with elevated levels of calcium and alkaline phosphatase. She had also sometimes exhibited slight elevation of parathyroid hormone (97 pg/ml) while receiving treatment for hypertension and heart failure with atrial fibrillation, which had been detected by the Department of Cardiology at 71 years of age. She was admitted with elevated blood glucose (422 mg/dl) and hemoglobin A_1c (12%) levels, as well as for further assessment of hyperparathyroidism. Her diabetes was treated with intensive insulin therapy (insulin lispro at 24 U per day). Assessment of hypercalcemia revealed that urinary calcium excretion was extremely low (0 mg/day) and the fractional excretion of Ca (FE_Ca) was 0%. Familial hypocalciuric hypercalcemia was suspected. No abnormalities of the parathyroid gland were found by ultrasonography and ^99mTc+Tl scintigraphy. The patient and her family members did not have any mutations in the extracellular domain or transmembrane domain of the calcium-sensing receptor, which has been reported to play an important role in Ca binding and intracellular signaling. However, the patient did have a missense mutation of codon 990 (R990G) in the intracellular domain, which has been reported to be a site of polymorphism, as well as a mutation of intron 6 (16 base pairs downstream from the exon-intron boundary: IVS6+16). One year later, she developed Grave’s disease that presented with fatigue and weight loss.

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We report a rare case of ruptured mycotic thoracic aortic aneurysm that required almost one month for correct diagnosis. A 71-year-old woman who had hemoptysis for several weeks was initially suspected to have lung cancer based on several examinations, including fluorine-18 fluorodeoxyglucose-positron emission tomography. However, contrast-enhanced computed tomography revealed a ruptured mycotic thoracic aortic aneurysm in the lower lobe of the left lung. She underwent emergency surgery carried out collaboratively by cardiovascular and general thoracic surgeons. En bloc resection of the aneurysm with the left lower lobe and in situ graft replacement of the descending aorta were performed successfully, although left lower lobectomy was difficult due to the insufficient segmentation of the upper and lower lobes and strong adherence of the aneurysm to the left lung. The clinical course was uneventful. The reason for survival for one month was thought to be that the rupture was covered by the lung. Because the resection of the lung is often difficult in cases in which the aneurysmal rupture shows extensive lung invasion, collaboration with cardiovascular and general thoracic surgeons is important.

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Observing vocal fold vibration is essential when investigating the causes of hoarseness of voice and determining the appropriate treatment. However, in Japan, it is difficult at present to obtain commercial high-speed digital imaging (HSDI) systems for this purpose because of a lack of marketing routes. Accordingly, we devised an HSDI system using a high-speed camera (monochrome HiSpec1). We chose this camera as it has been an established instrument in industrial and laboratory use; it also provided excellent light sensitivity, and used a 400-watt xenon light source for the acquisition of bright and clear images. The HSDI system enabled video capture of up to 4.4 s with our usual setting of 422 × 384 pixels at 3000 frames per second (fps). It can also capture videos in various settings such as 396 × 256 pixels at 4000 fps. In addition, we equipped the system with devices for concurrent audio waveform acquisition. Our novel laryngeal HSDI system could be a useful clinical tool for vocal fold examination in patients with voice problems when access to commercially available devices is limited.

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