Site-specific bone loss in senescence-accelerated mouse (SAMP6): a murine model for senile osteoporosis.

The senescence-accelerated mouse strain P6 (SAMP6) is a model of senile osteoporosis, which possesses many features of senile osteoporosis in humans. So far, little is known about the systemic bone microstructural changes that occur at multiple skeletal sites. In this study, we therefore, investigated site (vertebra, femur and tibia) dependence of bone microstructure and bone mineral density (BMD) in SAMP6 and the normal control mouse (SAMR1) at 5 and 12months of age using quantitative micro computed tomography (micro-CT) and image analysis software. As compared with SAMR1, the most prominent change in SAMP6 was the reduction of vertebral trabecular bone volume fraction (BV/TV) and trabecular BMD. Moderate decrease of trabecular bone mass was observed in the proximal tibia and distal femur. Increased marrow area and periosteal perimeter were investigated, though the cortical area and cortical thickness had no marked changes in the mid-tibial and mid-femoral cortical bones. These results indicate that bone microstructural properties in SAMP6 are remarkably heterogeneous throughout the skeleton, which is analogous to changes that occur in human bones. These findings further validate the relevance of SAMP6 as a model of senile osteoporosis.

Anatomic variation of the celiac trunk with special reference to hepatic artery patterns.

Based on a large homogeneous sample from a Japanese population, anatomic variations in the celiac trunk (CT) and the hepatic artery were studied. Previously we analyzed the branching mode of the CT in 450 Japanese cadavers. In order to maximize the database on the CT and hepatic artery, we examined the anatomy of these arteries in 524 cadavers, a total of 974 cases. A total of 89.8% of cases showed the classical trifurcation of the CT. The typical normal pattern of the CT and the hepatic artery was confirmed in 66.6% and 72.4% of the cadavers, respectively. Variant left and right hepatic arteries were observed in 11.0% and 4.9%, respectively. A variant anatomy involving both the left and right hepatic arteries was found in 1.5%. A common hepatosplenic trunk and a gastrohepatic trunk were seen in 4.4% and 0.3%, respectively. A common hepatic artery (CHA) arising from the superior mesenteric artery (SMA) or directly from the aorta was present in 3.5% or 0.5%, respectively. A hepatosplenomesenteric trunk and a celiomesenteric trunk were encountered in 0.7%, respectively. This anatomical update of the CT and the hepatic artery can be useful for transplantation and general surgeons, as well as vascular radiologists in this area.

Tibetan Medicated-Bath Therapy may Improve Adjuvant Arthritis in Rat.

Tibetan medicated-bath therapy has been applied to patients with rheumatoid arthritis for centuries. However, the detailed action mechanism of Tibetan medicated-bath therapy on the morphology and function of joints remains unknown. We designed our investigation to evaluate the efficacy of Tibetan medicated-bath therapy on adjuvant arthritis (AA) of rats in comparison with water-bath and dexamethasone administration. AA was induced by intradermal injection of Mycobacterium butyricum suspended in sterile mineral oil. The control animals were similarly injected with sterile vehicle. Eight days after injection, rats were treated with fresh-water bath, Tibetan medicated-bath (40 degrees C, 15 min) or intramuscular injection with dexamethasone for 21 consecutive days after which we evaluated the severity of arthritis visually and microscopically and measured serum interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha levels. While arthritis did not significantly change after water-bath treatment, the Tibetan medicated-bath and dexamethasone groups showed diminished joint swelling and alleviation of, inflammatory cell infiltration and the destruction of bone and cartilage. Serum IL-6 and TNF-alpha levels significantly decreased. Our results demonstrated that Tibetan medicated-bath therapy exerted a reliable effect on rat adjuvant arthritis, which may be involved in the inflammatory cytokines, IL-6 and TNF-alpha. Our data provide evidence for clinical use of Tibetan-medicated bath therapy for arthritis patients.

Three-dimensional microstructure of the bone in a hamster model of senile osteoporosis.

Age-related bone loss, which is poorly characterized, is a major underlying cause of osteoporotic fractures in the elderly. In order to identify the morphological feature of age-related bone loss, we investigated sex and site (tibia, femur and vertebra) dependence of bone microstructure in aging hamsters from 3 to 24 months of age using micro-CT. In the proximal tibia and distal femur, trabecular bone volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th) and bone mineral density (BMD) increased to a maximum at 6 or 12 months and then declined progressively from 12 to 24 months of age. Trabecular separation (Tb.Sp), trabecular bone pattern factor (TBPf) and structure model index (SMI) increased with age. As compared with male hamsters, BV/TV and Tb.N were significantly lower in females at 18 and 24 months of age. Age-related decrease of trabecular BV/TV in the vertebral body was less than that of the femoral and tibial metaphyses. In the mid-femoral diaphysis, cortical bone area remained constant from 3 to 24 months of age. Cortical thickness decreased from 12 to 24 months and cortical BMD declined significantly from 18 to 24 months of age. These findings indicate that skeletal site and sex differences exist in hamster bone structure. Age-related bone changes in hamsters resemble those in humans. We conclude that hamster may be a useful model to study at least some aspects of bone loss during human aging.

Scanning electron microscopic study of the human auditory ossicles.

The human mallei, incudes and stapedes from 34 cadavers were examined using scanning electron microscope (SEM) to compare the bone surface type among different regions of auditory ossicles for males and females. On the malleus of both males and females, almost all of the surfaces showed a smooth fibrous appearance, characteristic of resting surface. Limited bone-forming or resorbing surfaces were identified on the malleus. As compared with the malleus, the percentage area of the resorbing surface and the vascular canal openings were higher on the incus and stapes, especially on the long process (Crus longum) of the incus and the neck of the stapes for both males and females. The percentage area occupied by the resorbing surface of the long process of the incus and the neck of the stapes correlated with that of the vascular canal openings. We consider that the malleus maintained the stable condition, while the long process of the incus and the neck of the stapes demonstrated marked bone resorption. We suppose that the bone erosion may be related to the vascularization in these regions. Though the percentage area of the resorbing surface and the vascular canal openings had the tendency to be high in females, we did not find any significant differences between the males and females. There was no significant correlation between the age and the area of resorbing surface or vascular canal openings.

Morphologic study of the lateral and medial collateral ligaments of the human knee.

The collateral ligaments of the knee are important in maintaining knee stability. Numerous studies have been described the morphology and function of the ligaments of the knee. However, little data was given about the lateral and medial collateral ligaments simultaneously. The objective of the present study was to compare the morphology of the lateral (LCL) and medial (MCL) collateral ligaments in human knees. We dissected 34 cadaver knees and examined the size, attachment and inclination of the LCL and the MCL. In both of them, the posterior bundle was longer than that of the anterior bundle. The anterior and posterior bundles of the MCL were longer than that of the LCL. The proximal attachment of the MCL situated more anterior and superior than that of the LCL. As compared with the LCL, the distal attachment of the MCL located a little forward and inferior. The longitudinal diameter of the attachment of ligaments was longer than the transverse one, except for the proximal attachment of the MCL. The size of the attachments of the MCL was larger than that of the LCL except for the longitudinal diameter of the proximal attachment. At full extension of the knee, the inclination of the MCL was backward, while that of the LCL was forward. These data showed that the LCL and MCL had the different structural properties. It may provide a better understanding of the function of the collateral ligaments.

Effects of hPTH (1-34) and Gosha-jinki-gan on the trabecular bone microarchitecture in ovariectomized rat tibia.

Although human parathyroid hormone (1-34) [hPTH (1-34)] was reported to improve osteoporotic bone loss, little is known about the anti-osteoporotic effect of the traditional Chinese medicine, Gosha-jinki-gan (GJG). The purpose of this present study was to clarify and compare the effects of hPTH (1-34) and GJG on trabecular bone microarchitecture in ovariectomized (OVX) rat tibia by using microcomputed tomography (micro-CT). Thirty 12-week-old Sprague-Dawley female rats were underwent ovariectomy (OVX) or sham operation. Four weeks later, the ovariectomized rats were further divided into OVX, OVX + PTH, and OVX + GJG groups. hPTH (1-34) was administered subcutaneously at a dose of 20 microg/kg, 3 times/week, and OVX + GJG group received 0.05% aqueous solution of GJG as the only drinking fluid for 8 weeks respectively. The three-dimensional (3D) trabecular microarchitecture of the bone in the proximal tibial metaphysis was evaluated by micro-CT. In the OVX + PTH group, trabecular bone volume (BV/ TV), number (Tb.N) and thickness (Tb.Th) were significantly increased, structure model index (SMI) and trabecular bone pattern factor (TBPf) decreased when compared with the OVX group. In comparison to the OVX group, BV/TV and Tb.N were significantly greater, while SMI and TBPf had no marked changes in the OVX + GJG group. These results suggest that the administration of hPTH (1-34) restore the trabecular bone volume and improve the microstuctural property as well, while GJG reduce the bone loss without affecting its microstructural property in ovariectomized rats.

Morphological alterations in the growth plate cartilage of ovariectomized mice.

The effects of ovariectomy on growth and estrogen receptor (ER) expression level in the epiphyseal growth plate in mice have been estimated by histomorphometry and immunohistochemistry. Twelve female ddY mice, 8-9 weeks of age, were subjected to bilateral ovariectomy and 12 others were sham operated. They were then killed 8 weeks later. Ovariectomy significantly increased the total thickness of the distal femoral and proximal tibial growth plate cartilage. Ovariectomy caused a 1.4-fold increase in the thickness of the proliferative layer in the distal growth plate of the femur and a 1.3-fold increase in the thickness of the proliferative layer in the proximal growth plate of the tibia. ERalpha and ERbeta immunoreactivity was detected in chondrocytes of the growth plate and the expression level of ERs in epiphyseal plates was increased in ovariectomized mice compared with controls. These data suggest that ERalpha and ERbeta are coexpressed in the growth plates of the mice and that the cartilage growth and the level of expression of ERs in these tissues are hormonally regulated.

Morphology of the lingual papillae in the raccoon dog and fox.

The dorsal lingual surfaces of the raccoon dogs (Nyctereutes procyonoides) and fox (Vulpes vulpes japonica) were examined by scanning electron microscopical (SEM) observations. The distribution and type of the lingual papillae found in the raccoon dog were similar to those in the fox. Filiform, fungiform, foliate and vallate papillae were observed. The filiform papillae were distributed over the entire dosal surface of the tongue. Each filiform papilla on the apical surface of the tongue had several pointed processes. The filiform papillae of the lingual body consisted of a main papilla and some secondary papillae. The fungiform papillae were present rounded bodies, and more densely distributed on the lingual apex. The foliate papillae were seen on the dorsolateral aspect of the tongue. The vallate papillae were located on both sides of the posterior end of the lingual body. Each papilla was surrounded by groove and crescent pad. On the periphery of the papillae, large conical papillae were observed.

Comparative effects of milk and soymilk on bone loss in adult ovariectomized osteoporosis rat.

The present study was designed to investigate the comparative effects of milk and soymilk on the bone morphology of ovariectomized rat model with postmenopausal osteoporosis. Fourty 12-week-old female Spraque-Dawley rats were randomly divided into the following 5 groups: intact control (control), sham-operated (sham), ovariectomized (OVX), ovariectomized and fed milk (milk), ovariectomized and fed soymilk (soymilk) groups. Rats of OVX, milk and soymilk groups were bilaterally ovariectomized. Milk or soymilk of 3 ml was administered by gavage via an intragastric tube per day for 6 weeks. The distal femoral metaphysis was studied morphologically, together with bone mineral density (BMD), serum parathyroid hormone (PTH) and estradiol levels. In the OVX group, the trabecular bone volume was significantly reduced. The percentage area of resorbing surface was high. In the soymilk group, the trabecular bone volume was 68% higher and the percentage area of resorbing surface was 18% lower as compared with the OVX group. The serum estradiol level rose 48% and the serum PTH level fell 26% in the soymilk group. In the milk group, the trabecular bone volume had the tendency to be high, but there was no significant change of the bone morphology. The present study provides certain evidence that the soymilk has beneficial effects in preventing bone loss in rats induced by ovarian hormone deficiency.

Bilateral thoracic ducts with coexistent persistent left superior vena cava.

A case of bilateral thoracic ducts with coexistent persistent left superior vena cava (SVC) was identified in a 77-year-old Japanese female cadaver during dissection in a gross anatomy course. The persistent left SVC began at the lower surface of the left brachiocephalic vein, descended in front of the aortic arch, and drained into the right atrium through the coronary sinus. The right SVC was normal both in size and in position. The azygos vein, receiving the hemiazygos vein, opened into the right SVC. The accessory hemiazygos vein and the left superior intercostal vein united to form a common trunk, which drained into the left SVC. The left and right thoracic ducts began at the level of the 1st lumbar vertebra, ran upwards parallel and anterior to the vertebral column, and terminated at the venous angles of their corresponding sides. There was an anastomotic branch between them. The present case was considered to be very rare, since the persistent left SVC and bilateral thoracic ducts coexisted. The embryologic basis and clinical importance of this case are discussed.

Effects of traditional Chinese medicine on bone loss in SAMP6: a murine model for senile osteoporosis.

We evaluated the effects of the traditional Chinese medicines, Hachimi-jio-gan, Juzen-taiho-to and Unkei-to, on bone loss in murine model of senile osteoporosis (SAMP6). Two-month-old SAMP6 were divided into control and experimental groups. The control mice had the tap water available as the only drinking fluid. The experimental mice were given 0.05% aqueous solution of Hachimi-jio-gan, Juzen-taiho-to or Unkei-to for three months. The solution intake of a mouse averaged 5 ml per day. The bones were studied morphologically and histomorphometrically, together with bone mineral density (BMD), serum parathyroid hormone (PTH) and estradiol levels. In the control group, BMD and the amount of bone forming surface were low, the serum PTH level was high when compared with the normal mice SAMR1. Many osteocytes and osteoblasts showed degenerative changes and numerous mast cells were observed in the bone marrow. Compared with controls, the serum estradiol level was higher in the Unkei-to group. However, we did not find any significant changes of bones. In the Hachimi-jio-gan and Juzen-taiho-to groups, the bone mass and the amount of bone forming surface increased. Most of the osteocytes and osteoblasts appeared normal. As compared with controls, the number of mast cells in bone marrow decreased in the Hachimi-jio-gan group. The serum PTH level had declined in the Juzen-taiho-to group. The present study provides certain evidence that Hachimi-jio-gan and Juzen-taiho-to are effective in preventing bone loss in SAMP6, while Unkei-to can only improve the ovary function.

Morphology of the lingual papillae in the tiger.

The dorsal lingual surfaces of an adult tiger (Panthera tigris altaica) was examined by macroscopical and scanning electron microscopical observations. Filiform, fungiform and vallate papillae were observed. The filiform papillae were distributed over the entire dosal surface of the tongue. The fungiform papillae were present rounded bodies, and more densely distributed on the lingual apex. There were 4 vallate papillae in total on borderline between the lingual body and lingual radix. Each papilla was surrounded by a groove. No foliate papillae were seen on the dorsal surface. Openings of the glandular ducts on the regions of the vallate papillae were found.

SEM study on the dorsal lingual surface of the large flying fox, Pteropus vampyrus.

The dorsal lingual surface of the large flying foxes were examined by scanning electron microscopy. Filiform, fungiform and vallate papillae were observed. The filiform papillae were distributed over the entire dorsal surface of the tongue. The filiform papillae notably differed in morphology by their location on the tongue and could be classified into 6 types: 1) scale-like, 2) giant trifid, 3) small crown-like, 4) large crown-like, 5) long conical and 6) short conical papillae. The fungiform papillae were present rounded bodies on the anterior 2/3 of the tongue. The large flying foxes showed the triangular arrangement of the three vallate papillae, with the apex of the triangle directed posteriorly. These findings indicate that the tongue of the large flying bat is similar to that of the lesser dog-faced fruit bat.

Ultrastructure of the parathyroid gland of the japanese lizard in the spring and summer season.

The ultrastructure of the parathyroid glands of adult Japanese lizards (Takydromus tachydromoides) in the spring and summer season was examined. The parenchyma of the gland consists of chief cells arranged in cords or solid masses. Many chief cells contain numerous free ribosomes and mitochondria, well-developed Golgi complexes, a few lysosome-like bodies, some multivesicular bodies and relatively numerous lipid droplets. The endoplasmic reticulum is mainly smooth-surfaced. Cisternae of the rough endoplasmic reticulum are distributed randomly in the cytoplasm. Small coated vesicles of 700-800 Å in diameter are found occasionally in the cytoplasm, especially in the Golgi region. The chief cells contain occasional secretory granules of 150-300 nm in diameter that are distributed randomly in the cytoplasm and lie close to the plasma membrane. Electron dense material similar to the contents of the secretory granules is observed in the enlarged intercellular space. These findings suggest that the secretory granules may be discharged into the intercellular space by an eruptocrine type of secretion. Coated vesicles (invaginations) connected to the plasma membrane and smooth vesicles arranged in a row near the plasma membrane are observed. It is suggested that such coated vesicles may take up extracellular proteins. The accumulation of microfilaments is sometimes recognized. Morphological evidence of synthetic and secretory activities in the chief cells suggests active parathyroid function in the Japanese lizard during the spring and summer season.