Chlorella extract （CE） has been shown to induce production of T helper-1 cytokines, and regulate serum IgE levels in animal models of asthma. We aimed to evaluate whether CE could inhibit ovalbumin （OVA）-induced airway hyperresponsiveness （AHR） and airway remodeling in a murine model of asthma. Balb/c mice were allocated to four groups: a control group （no OVA exposure, not given CE）, a CE group （no OVA exposure, given CE）, an asthma group （sensitized/challenged with OVA, not given CE） and a CE＋asthma group （sensitized/challenged with OVA, given CE）. In the asthma and CE＋asthma groups, mice were sensitized with OVA on day 0 and day 12, and then challenged with OVA on three consecutive days. In the CE and CE＋asthma groups, the mice were given feed containing 2％ CE. We assessed AHR to methacholine, and analyzed bronchoalveolar lavage fluid （BALF）, serum, lung tissue and spleen cells. Administration of CE was associated with significantly lower AHR in OVA-sensitized and challenged mice. CE administration was also associated with marked reduction of total cells, eosinophils and T helper-2 cytokines （IL-4, IL-5 and IL-13） in BALF. In addition, administration of CE significantly decreased the numbers of periodic acid-Schiff （PAS）-positive cells in OVA-sensitized and challenged mice. Administration of CE also directly suppressed IL-4, IL-5 and IL-13 production in spleen cells of OVA-sensitized and challenged mice. These results indicate that CE can partly prevent AHR and airway remodeling in a murine model of asthma.

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This study investigated the relationships of pre- to postoperative changes in posterior condylar offset（PCO）to differences in condylar length and range of motion in posterior stabilized（PS）-type total knee arthroplasty（TKA）. Subjects and Methods: We studied 40 knees of 40 patients（10 males and 30 females）treated by PS-type TKA. Slide calipers were used to measure condyle lengths intraoperatively and PCO was measured by standard X-ray. The patients were divided into two groups based on a pre- to postoperative change in PCO of ≥ 3mm or＜3mm, with the differences in both condylar length and range of motion compared between groups. The mean differences in condyle lengths were 2.6mm（33 knees）and 4.3mm（7 knees）in cases with PCO changes of＜3mm and ≥ 3mm, respectively. The means in the respective groups were −6° and −7° for preoperative extension, 116° and 118° for preoperative flexion, −3° and −4° for postoperative extension, and 131° and 129° for postoperative flexion. There was no significant difference in the range of motion between the groups. Cases with a large difference in condylar lengths were likely to have a small PCO postoperatively; however, the postoperative range of knee flexion was not significantly related to a small postoperative PCO. These findings suggested that preoperative range of knee motion, age, and type of TKA could influence both the postoperative range of motion and PCO.

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Neuroendocrine ductal carcinoma in situ of the breast（NE-DCIS）was recently recognized as a special subtype of DCIS, although the diagnostic criteria for NE-DCIS are yet to be established. DCIS is defined as the immunohistochemical expression of neuroendocrine markers chromogranin A and/or synaptophysin in over 50％ of tumor cells. Here, we investigated whether there are significant differences in magnetic resonance imaging（MRI）findings between NE-DCIS and non-NE-DCIS. The study sample comprised 8 lesions in 7 patients with breast NE-DCIS and 71 lesions in 69 patients with non-NE-DCIS who underwent preoperative MRI and histopathological diagnosis at our hospital from June 2010 to June 2012. The patients were females aged 34–85 years. We examined the lesion type, pattern of time-signal intensity curve（TIC）on dynamic contrast-enhanced MRI（DCE-MRI）, presence or absence of bloody duct ectasia delineation, and presence or absence of calcification on mammography（MMG）. Mass-type lesions were significantly more common in breast NE-DCIS than in non-NE-DCIS on MRI. On DCE-MRI, the TIC washout pattern was more commonly observed in NE-DCIS than in non-NE-DCIS, and although there was no significant difference in the rate of bloody duct ectasia delineation, it was relatively more common in NE-DCIS. MMG revealed a significant difference in calcification between non-NE-DCIS（60.1％）and NE-DCIS（0％）. Mass-type lesions and TIC washout pattern are significantly more common in patients with NE-DCIS than in those with non-NE-DCIS on MRI and DCE-MRI.

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Polypodium leucotomos（PL）is a fern native to Latin America, and its extract is used as an oral sunscreen; however, its safety during use has not been adequately investigated. Therefore, the aim of this study was to evaluate food-drug interactions associated with PL extract mediated by cytochrome P450 3A（CYP3A）inhibition and induction. Inhibition of CYP3A-mediated midazolam（MDZ）1’-hydroxylation activity by PL extract and its major phenolic components was evaluated in vitro using pooled human liver microsomes. In addition, MDZ pharmacokinetics were investigated in rats after a single dose, as well as after 1 week treatment with PL extract（30mg/kg）in order to evaluate the inhibitory and inducible effects of PL on CYP3A in vivo, respectively. Serum MDZ concentrations were analyzed and pharmacokinetic parameters were compared between PL- and water（control）-treated groups. In vitro, PL extract decreased MDZ 1’-hydroxylation activity in a concentration-dependent manner. However, the major phenolic compounds in PL extracts, namely caffeic, chlorogenic, p-coumaric, ferulic, and vanillic acids, did not exhibit any marked inhibitory effects on MDZ 1’-hydroxylation activity. In vivo, administration of a single dose of PL extract to rats significantly increased the area under the serum concentration-time curve from time 0 to infinity（AUC_0–∞）and the maximum serum concentration（C_max）of MDZ（by 57％ and 88％, respectively; P＜0.05）. In contrast, there were no significant changes in MDZ pharmacokinetic parameters after 1 week of treatment with PL extract. These results suggest that PL extract can cause a food-drug interaction by inhibiting CYP3A.

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Gene silencing by RNA interference （RNAi） is widely used for assessing gene function. An important advance in the RNAi field was the discovery that plasmid-based RNAi can substitute for synthetic small interfering RNA in vitro and in vivo. However, constitutive and ubiquitous knockdown of gene expression by RNAi in mice can limit the scope of experiments because this process can lead to embryonic lethality, or result in compensatory overexpression of other genes such that no phenotypic abnormalities occur. Either way, analyses of the physiological roles of the gene of interest in adult mice are not possible. To overcome these limitations, we previously constructed a double-conditional short-hairpin RNA （shRNA） expression vector that can regulate shRNA expression in a spatio-temporal manner with a tetracycline-inducible floxed stuffer sequence selectively excised by application of Cre recombinase. In this study, we aimed to modify this vector to create an all-in-one vector that produces double-conditional transgenic mice through a single round of gene transfer to fertilized eggs. We added a coding region for nuclear localizing Cre （NCre） recombinase with a multi-cloning site for a cell-specific promoter into the double-conditional short-hairpin RNA （shRNA） expression vector that we previously constructed. Using Escherichia coli, we confirmed successful construction of the vector. First, we confirmed isopropyl-β-D-thiogalactopyranoside-induced expression of NCre recombinase through the lac operon as a specific promoter by western blotting. Second, we confirmed functional recombination of the floxed sequence of loxP-like TATA-lox by analysing restriction enzyme-digested fragments. This all-in-one double-conditional shRNA expression vector will be useful for reversible in vitro and in vivo knockdown of target gene expression, in target cells via promoter-specific expression of NCre, and at specific times by tetracycline application.

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Osteoarthritis is a complicated clinical condition affected by age, mechanical stress, cartilage hypertrophy, cytokines, and genetic predisposition. In this study, we compared the effects of various anti-inflammatory analgesics on mechanical stress-induced inflammation in a synovial sarcoma cell line （SW982 cells）. SW982 cells exposed to mechanical stress by shaking with hydroxyapatite-simulating bone chips were treated with acetaminophen, ketoprofen, triamcinolone acetonide, celecoxib, or neurotrophin for 48hr. The expression of integrin α5β1 receptor, observed in fibroblasts and synovium, was evaluated. Levels of the transcription factor, nuclear factor-κB, the inflammatory cytokine, tumor necrosis factor-α, the proteolytic enzyme, matrix metalloproteinase-3, and prostaglandin E2, which is associated with pain and arachidonate cascade product levels, were measured by ELISA. The expression of integrin α5β1 was significantly increased by mechanical stress. Activation of nuclear factor-κB by mechanical stress was significantly suppressed by celecoxib only. Mechanical stress-induced increases in tumor necrosis factor-α and matrix metalloproteinase-3 levels were significantly suppressed by acetaminophen, triamcinolone acetonide, and neurotrophin. The mechanical stress-induced increase in prostaglandin E2 levels was significantly suppressed by acetaminophen, ketoprofen, and celecoxib. SW982 exposed to mechanical stress is proposed as a model for arthritis, and indeed, the expression of integrin α5β1, a membrane receptor protein that binds to fibronectin and the extracellular matrix, and is involved in cell proliferation, differentiation, and neovascularization in osteoarthritis, was significantly upregulated. Following evaluation using this model, acetaminophen was found to possess anti-inflammatory, analgesic, and joint-destruction suppression properties. This drug may, therefore, have applications in the treatment of mechanical stress-induced inflammation.

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Despite many reports describing the surgical procedure for knee ligament reconstruction, especially the anterior cruciate ligament, there have been few reports on infection prevention and none on sterilizing reconstruction materials using povidone-iodine during surgery. Here we report our surgical techniques for infection prevention. The participants included 522 patients who underwent arthroscopic knee ligament reconstruction at our hospital from April 2001 to October 2005. Despite taking various measures after cases of infection in 2001, such as the selection of sutures and artificial ligaments, and the use of indwelling intra-articular drains, the infection rate was not reduced to zero. Recently, we felt that soaking reconstruction ligaments in povidone-iodine and then washing with saline （“povidone-iodine pickling”） immediately prior to burr-hole insertion may prevent bacterial infections caused by surgical instruments and the surgery. Therefore, in this study, we examined the effects of our techniques for intraoperative and postoperative infection prevention, including “povidone-iodine pickling”. Although there were 6 cases of infection out of 100 in 2001, 3 out of 112 in 2002, 2 out of 121 in 2003, and 1 out of 121 in 2004, there were no cases of infection from September 2004 to October 2005 when “povidone-iodine pickling” was positively carried out. According to a 2015 study on the use of povidone-iodine, immersing a graft dropped on the floor during surgery in a chlorhexidine gluconate or povidone-iodine solution significantly reduced contamination of the graft. While the use of povidone-iodine for infection prevention may produce cytotoxicity, it is essential to mitigate the risk of septic arthritis. Going forward, we need to understand the specific pathologic basis of any cytotoxicity caused by the use of povidone-iodine on reconstructed ligaments.

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An 82-year-old woman had been taking alendronate for 5 years and 5 months, which had been prescribed for osteoporosis at a local clinic. Severe left thigh pain began without any trauma. X-ray, computed tomography and magnetic resonance imaging findings showed atypical femoral fracture（AFF）. Treatment with teriparatide and weight-bearing therapy was selected. Bone union was achieved without surgery. Teriparatide has been reported to promote AFF healing. At four years and 9 months from the beginning of treatment, our patient has no left femoral pain and no impairment to activities of daily living. Careful follow-up will be necessary as long-term outcomes of conservative AFF treatment have not been reported to date.

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Acute pancreatitis（AP）in childhood is rare, but is starting to be recognized more often. However, optimal enteral nutrition formulas for children with AP have not been determined. This report describes the successful management of AP, with low-fat elemental enteral nutrition, in a 5-year-old boy. The patient had been diagnosed with mild AP and started oral feeding after about 3 days of fasting, but his pancreatic enzyme levels subsequently became elevated. Therefore, to allow the pancreas to rest, total parenteral nutrition（TPN）was started. After starting TPN, there was no improvement in his pancreatic enzyme levels but, because his clinical symptoms were stable, a small amount of elemental diet was initiated. Elemental diet therapy was safely performed without return of any clinical symptoms and the patient’s pancreatic enzyme levels slowly improved. Elemental diet was both safe and beneficial, providing clinical remission and improvement in quality of life. Early elemental diet therapy may therefore be a useful treatment strategy for pediatric patients with AP.

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