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Abnormal hip bone morphologies are associated with various diseases of the hip joint. Weight bearing, especially during growth, may be important to achieve normal acetabulum development. This study aimed to investigate whether hip bone morphologies were affected by hindlimb suspension (HS) in 4 week-old rats. In HS groups, tail suspension was applied for 0, 2, 4, and 8 weeks. Age-matched rats were used as controls. The complex of hip bones with lumbar and sacral vertebrae were assessed based on morphological indexes using three-dimensional reconstructed images from X-ray computed tomography. Acetabular widths (measured from cranial to caudal) unchanged and depths became larger in both groups with age. Acetabular lengths (from the ventral side to the dorsal side) became larger in control groups but unchanged in HS groups with age. In HS groups, acetabular width, length, and depths were smaller than the control groups at 4 and/or 8 weeks. Acetabular versions became enlarged (rotated inwards) with age in both groups, although this was particularly pronounced in HS groups. Histologically, triradiate cartilage layers in the acetabulum were thinner with age and almost disappeared at 8 weeks in both groups. However, HS decreased Safranin O staining and prolonged the presence of hypertrophic chondrocyte indicating alterations in the chondral ossification processes. Iliac wing angles remained unchanged and anterior superior iliac crest (ASIC) distances increased with age in controls. In contrast, HS groups showed narrowed iliac wing angles with small ASIC distances. These results suggest that reduced mechanical loading during growth can interfere with hip joint formation. Keywords Hindlimb suspension, Hip joint, Acetabular morphology, Triradiate cartilage.

The Asian cultivated rice, Oryza sativa, is one of the most important crops feeding more than a third of global population. In spite of the studies for several decades, the origin and domestication history of rice varietal groups, japonica and indica, have not been fully unveiled. Genetic information of ancient rice remains is essential for direct and exclusive insight into the domestication history of rice. We performed ancient DNA analysis of 950- to 2,800-year-old rice remains excavated from Japan and Korea. We found the presence of both japonica- and indica-type varieties in the Yayoi period and the middle ages of Japan and the middle part of Korea Peninsula 2,000 years ago. It is popularly considered that japonica has been exclusively cultivated in northern part of East Asia including Japan and Korea. Our result disclosed unexpectedly wide diversity of rice varieties in archaic East Asia. The present results from ancient rice DNA reveal an exclusive insight for the domestication history of rice which is not provided as far as contemporary rice., (© The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Synthesis of ω-alkenyl-terminated silver nanoparticles (AgNPs) and then their immobilization on a hydrogen-terminated silicon surface in two-dimensional arrangement through covalent interaction are demonstrated. The thermal-induced hydrosilylation at mild conditions facilitate nanoparticles assembly through interaction between terminal alkenyl (CH(2)=CH-) groups of AgNPs and hydrogen-terminated silicon surface. The assembly of AgNPs on a silicon surface is characterized by FESEM and XPS. Adequate coating of 10-undecene-1-thiol (UDT) on AgNPs and mild temperature hydrosilylation impede the fusion or aggregation of nanoparticles, while they immobilized on a silicon surface, which is very crucial to preserve the discrete entities of nanoparticles. This elegant and facile approach provides stable monolayer of AgNPs with very good coverage area and promises potential to fabricate electronic devices and solar cells, where nanoparticles needs to be directly attached to the silicon surface without an interfacial oxide thin film., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Recent studies suggest that plasminogen activator inhibitor-1 (PAI-1), a major inhibitor of the fibrinolytic system, may promote the development of asthma. To further investigate the significance of PAI-1 in the pathogenesis of asthma and determine the possibility that PAI-1 could be a therapeutic target for asthma, this study was conducted. First, PAI-1 levels in induced sputum (IS) from asthmatic subjects and healthy controls were measured. In asthmatic subjects, IS PAI-1 levels were elevated, compared with that of healthy controls, and were significantly higher in patients with long-duration asthma compared with short-duration asthma. PAI-1 levels were also found to correlate with IS transforming growth factor-β levels. Then, acute and chronic asthma models induced by ovalbumin were established in PAI-1-deficient mice and wild-type mice that received intra-airway administrations of small interfering RNA against PAI-1 (PAI-1-siRNA). We could demonstrate that eosinophilic airway inflammation and airway hyperresponsiveness were reduced in an acute asthma model, and airway remodeling was suppressed in a chronic asthma model in both PAI-1-deficient mice and wild-type mice that received intra-airway administration of PAI-1-siRNA. These results indicate that PAI-1 is strongly involved in the pathogenesis of asthma, and intra-airway administration of PAI-1-siRNA may be able to become a new therapeutic approach for asthma.

S-1 is a novel oral fluorouracil prodrug that plays a role in non-small cell lung cancer (NSCLC). Docetaxel (DTX) is one of the standard agents for relapsed NSCLC. We performed a phase I study of DTX plus S-1 combination therapy as second-line treatment for NSCLC to determine the maximum tolerated dose (MTD) and recommended dose (RD). Patients with recurrent NSCLC, aged 20-74 years with an Eastern Cooperative Oncology Group performance status of 0-1 and measurable lesions, were enrolled. The treatment consisted of four dose levels. The patients received DTX (40-60 mg/m(2) intravenously on day 1) and S-1 (65-80 mg/m(2) orally, daily on days 1-14) for each 21-day cycle. Three to six patients were treated at each dose level with the two drugs, with MTD defined as the dose level at which dose-limiting toxicity (DLT) occurred in 33% of the patients. A total of 17 patients were enrolled. At dose level 4 (DTX, 60 mg/m(2); S-1, 80 mg/m(2)) 3 of 5 patients experienced DLT and this level was regarded as the MTD. Therefore, dose level 3 (DTX, 60 mg/m(2); S-1, 65 mg/m(2)) was selected as the RD for subsequent studies. The DLTs were neutropenia (grade 4) and mucositis (grade 3). The response rate was 5.9% (1 of 17 patients achieved a partial response) and 14 of 17 patients achieved stable disease. This combination regimen showed a tolerable and manageable profile in recurrent NSCLC and therefore warrants further evaluation.

Objective: This study was originally designed to observe the effects of propranolol (a beta-blocker) and Zhigancao Decoction ([Chinese characters: see test] ZGCD) on bone mass in ovariectomized rats., Methods: Thirty-eight female Sprague-Dawley rats were divided into four groups initially, a sham-operated group (Sham, n=7), a model ovariectomized (OVX) group (Model, n=7), a propranolol group (Pro, n=12) and a ZGCD group (ZGCD, n=12). After 15 weeks of treatment, the expected effects were not found. In order to verify the situations of the experiment, we modified the study by administering calcitonin to a subgroup of the tested Pro and ZGCD rats., Results: The Pro and ZGCD treatments showed decreased heart rate and plasma norepinephrine level, but neither an increased bone mass nor any bone metabolism differences from the model rats were found. However, the OVX-induced bone loss was prevented by the sequent treatment of calcitonin., Conclusions: The results provide no evidence that the beta-blocker propranolol may stimulate bone formation, and do not justify its use for clinical treatment of osteoporosis.

Adenine is widely used in clinical field, however, an excess of adenine is harmful. It is known that the feeding of an adenine-rich diet induces renal failure and decreases bone mineral density (BMD) and the serum testosterone level in male rats. However, there is little information about the influence of adenine on female animals. We compared the effects of adenine treatment between male and female rats. Young male and female rats were administered adenine adjusted with distilled water (6 mg/ml, 50 mg/ml and 100 mg/ml) for 8 weeks (3 times/week, 8-16 week old). In male rats, renal failure was induced by 100 mg/ml adenine treatment and renal dysfunction was induced by 50 mg/ml adenine treatment. Bone loss and the reduction of the testosterone level were also caused by both concentrations of adenine. However, the serum testosterone level and BMD in male rats were decreased by 6 mg/ml adenine treatment by which renal dysfunction was not caused. It is suggested that adenine directly affected bone metabolism and sex steroidgenesis in male animals, not through altering renal dysfunction. In female rats, conversely, renal dysfunction was induced only in the 100 mg/ml group, which was somewhat different from the observation in male rats. The serum 17-beta estradiol level and the BMD in female rats were not affected by adenine treatment at all. In conclusion, there is a significant difference of the effects of adenine, which is commonly contained in medicine and general foods, on steroidgenesis and renal function between male and female rats.

The effects of acupuncture on bone biomechanical properties and histomorphometry in ovariectomized (OVX) rats were studied. Twenty-four 8-week-old female Sprague-Dawley rats were randomly assigned to three groups: sham, model and acupuncture. Rats in the model and acupuncture groups were ovariectomized, while those in the sham group underwent a sham operation. All rats were anesthetized and fastened for 15 minutes, and for the acupuncture group, needling on Pishu (BL20) and Shenshu (BL23) was performed. Blood and urine were collected to measure serum osteocalcin (OC) and urinary calcium, phosphorus or deoxypyridinoline (Dpd). After 16 weeks of treatment, all the rats were killed and their tibiae and femora were removed. The tibiae were used for analyses of bone histomorphometry and the femora for a three-point bending test. Acupuncture gave significant protection against ovariectomy-caused decline on femoral strength in the mechanical test, increased the trabecular bone volume and thickness, lowered the trabecular separation of tibiae and restricted the excretion of phosphorus and Dpd, while promoting the concentrations of serum osteocalcin as compared with model rats. These results seemed to indicate that acupuncture on the points of Pishu (BL20) and Shenshu (BL23) not only promoted the bone formation but also suppressed the bone resorption induced by OVX in osteoporotic rats, which suggests that it would be a potentially useful and convenient method in preventing osteoporosis.

The effects of Chinese herbal medicines including Hachimi-jio-gan (HJG) and/or Hochu-ekki-to (HEW) on osteopenia in rats were investigated. The Chinese herbal medicines were administrated for 8 weeks (7 times/week) starting from 1 week after ovariectomy. HJG and Prescription-2 (Prsc-2, the prescription based on HJG) showed protective effect on bone loss of the vertebrae after ovariectomy. However, Prescription-1 (the prescription based on HJG and HET) had no effect. Then, we made osteopenia model in rats by prednisolone and low calcium diet. Prsc-2 and HJG were administrated for 20 weeks with prednisolone. These Chinese remedies showed protective effects for osteopenia, with better indices on bone loss of the limbs than HJG alone in the osteopenia rats. It can be concluded that Prsc-2 is more effective than HJG for bone loss induced by various factors, and the additives in Prsc-2 may enhance the therapeutic effect.

In the adenine-induced renal failure rats, reversibility of renal failure and recovery of bone mineral density (BMD) by discontinuation of adenine-rich diet were reported. We think that the effect to bone metabolism with medication may be able to be evaluated as reinforcement of the BMD recovery. We have so far investigated the Chinese herbal medicine based on Hachimi-jio-gan (HJG) which are more effective than HJG alone. In this study, we investigated the effects of our Chinese herbal prescription on BMD in the adenine-treated rats compared to that of vitamin D3 treatment. Young male rats were administrated 100 mg/ml adenine for 8 weeks, and they showed renal failure and bone loss. The adenine-treated rats were divided into the following 3 groups, that is, the group experienced no treatment (control), the group given our Chinese herbal medicine (HAO), and the group given vitamin D3 (VD3) medication. It is likely that VD3 medication was less effective for increase of the femoral BMD than increase of the spinal BMD. In contrast, HAO was effective for increase of the femoral BMD. The VD3 group showed low deoxypyridinoline (Dpd: bone resorption maker) as compared to the control group. However, the HAO group showed same or slightly high Dpd. It is suggested that VD3 may increase BMD by reduction of bone resorption, while HAO may show effect on BMD by activating bone metabolism. It is indicated that HAO may become a curative medicine for bone loss because of the different target site from vitamin D3.

In the silkworm, Bombyx mori, nonsusceptibility to B. mori densonucleosis virus type-2 (BmDNV-2) is controlled by a recessive gene, nsd-2 (nonsusceptibility to DNV-2). We investigated the genetic linkage between two random-amplified polymorphic DNA (RAPD) markers and the +nsd-2 gene. Initially, we constructed the JSD-2 strain (nsd-2/+), which is congenic to strain J137 (nsd-2/nsd-2) with respect to the chromosome containing the +nsd-2 gene, starting with a female of strain J137 and a male of strain C137 (+nsd-2/+nsd-2). Genomic DNAs were compared between infected individuals of the JSD-2 strain and J137 by a polymerase chain reaction (PCR) with 700 arbitrary 10-mer primers. Two RAPD markers (OPH19R and OPP01R) linked to the +nsd-2 gene were found. For the crossing-over experiment, a female of J137 was crossed with a male (nsd-2/+) of JSD-2. Segregation analysis showed that the most closely linked RAPD marker (OPP01R) was mapped 4.7 cM distant from +nsd-2.

Genomic DNAs were compared between males and females of the domesticated silkworm, Bombyx mori, strains C108, C137, J137, p50, and WILD-W (constructed by crossing a wild silkworm, B. mandarina, female with a male of strain C108) by polymerase chain reaction (PCR) with 700 arbitrary 10-mer primers. Four female-specific RAPDs (W-Kabuki, W-Samurai, W-Kamikaze, and W-Yamato) were found. The sex chromosome formulas of B. mori and B. mandarina are ZW (XY) for the female and ZZ (XX) for the male. The four female-specific RAPDs are assumed to be derived from the W chromosome because the other chromosomes are shared by both sexes. A computer search for deduced amino acid sequences of these four RAPDs revealed that all of them showed homology to previously reported amino acid sequences encoded in known retrotransposable elements from various organisms.

In the silkworm, Bombyx mori, nonsusceptibility to B. mori densonucleosis virus type-1 (BmDNV-1) is controlled by a recessive gene, nsd-1 (nonsusceptibility to DNV-1), located on the twenty-first chromosome. We investigated genetic linkage between five random amplified polymorphic DNA (RAPD) markers and the +nsd-1 gene. Initially, we constructed the CSD-1 strain (nsd-1/+) which is congenic to strain C137 (nsd-1/nsd-1) for the twenty-first chromosome, starting with a female of C137 and a male of strain J137 (+nsd-1/+nsd-1). For the crossing over experiment, a female of C137 was crossed with a male (nsd-1/+) of CSD-1. Segregation analysis showed that the most closely linked RAPD marker mapped 3.0 cM distant from +nsd-1. A more specific marker for +nsd-1 was made by converting this RAPD band into a sequence characterized amplified region (SCAR) using a series of newly designed primer pairs based on its DNA sequence.