Expression of neurotransmitters, vasculogenesis markers and myosin heavy chain isoforms in the masseter muscle of senescence-accelerated mouse prone 8 mice.

BACKGROUND: Learning and memory deficits and pathologic changes in the hippocampus caused by toothlessness and soft diet feeding are related to reduced masseter muscle (MM) function. OBJECTIVE: Myosin heavy chain (MyHC) isoform expression in the MM also changes under different chewing conditions. The neurotransmitter calcitonin gene-related peptide (CGRP) and vascular endothelial growth factor A (VEGF-A) are involved in MM formation. However, the relationship between CGRP, VEGF-A and MyHC isoforms in the MM in the senescence-accelerated mouse prone 8 (SAMP8) strain, a model of learning and memory deficits, remains unclear. METHODS: Changes in CGRP, VEGF-A, vasculogenesis marker and MyHC isoform mRNA expression in the MMs of ageing SAMP8 and senescence-accelerated mouse resistant 1 (SAMR1) mice was investigated through quantitative real-time polymerase chain reaction (qRT-PCR) and in situ hybridization. RESULTS: qRT-PCR revealed obviously high CGRP levels in the SAMP8 mouse MM (p < .001). MyHC-IId/x mRNA expression in the MM was higher in 24-week-old SAMP8 mice than 24-week-old SAMR1 mice (p < .001) but lower in slow-MyHC SAMP8 mice than SAMR1 mice (p < .001). CGRP mRNA was observed on the muscle fibres of the SAMP8 mouse MM but not the SAMR1 mouse MM through in situ hybridization. Principal component analysis (PCA) revealed strong positive contributions of SAMP8-MyHC-IId/x, SAMP8-CGRP, SAMR1-MyHC-emb, SAMR1-CGRP, SAMR1-VEGF-A, SAMR1-CD31, SAMP8-VEGF-A, and SAMP8-CD31 in the MM at 12 and 24 weeks. CONCLUSION: Calcitonin gene-related peptide is also key for the MyHC-IId/x and slow-MyHC patterns in the MMs of SAMP8 mice.

Structural and CBCT analysis of mandibular canal microvessels expressing neurotransmitters in human cadavers.

PURPOSE: This study focused on the detailed structure of microvessels of the neurotransmitter-positive vasa nervorum of the inferior alveolar nerve, vein, and artery in the mandibular canal (MC) to obtain information for improved safety in dental treatments. We also observed the detailed structure of the MC from the mental foramen to the mandibular foramen using cone-beam computed tomography (CBCT). METHODS: In this study, mandibles from 45 sides of 23 human cadavers aged 76-104 years were examined by microscopy, immunohistochemistry, and CBCT analysis. These data were further evaluated by principal component analysis (PCA). RESULTS: The microvessels of the vasa nervorum with calcitonin gene-related peptide- and neuropeptide Y-positive reactions were classified into 5 types: large (4.19%, 28/667); irregular large (7.35%, 49/667), numerous intermediate (29.23%, 195/667), irregular intermediate (29.23%, 195/667), and scattered fine (30.0%, 200/667) microvessels. The MC showed various structures from the 3rd molar to the premolars and was also classified into three types, including complete (57.0%, 228/400), partial (33.8%, 135/400), and unclear (9.2%, 37/400), from the mandibular foramen to the mental foramen. PCA results revealed that developed capillaries were mainly localized in the molar region. CONCLUSIONS: Fine microvessels of the vasa nervorum expressing neurotransmitters are present from the molar to premolar region, which is key information for mandibular dental treatments. The different microvessel structures also indicate differences in specific characteristics between dentulous and edentulous cadavers regarding oral surgical and implant treatments.

Relationship between the bilateral ratios of the thoracic shape and electromyographic activity of the thoracic and lumbar iliocostalis muscles during thoracic lateral translation.

[Purpose] This study aimed to determine the relationship between thoracic lateral deviation, the bilateral ratio of the thoracic shape, and the bilateral ratio of the thoracic and lumbar iliocostalis muscles during resting sitting and thoracic lateral translation. [Participants and Methods] We included 23 healthy adult males in the study. The measurement tasks were resting sitting and thoracic lateral translation relative to the pelvis. The thoracic lateral deviation and bilateral ratio of the upper and lower thoracic shapes were measured using three-dimensional motion capture. The bilateral ratio of the thoracic and lumbar iliocostalis muscles were measured using the surface electromyographic recording. [Results] The bilateral ratio of the lower thoracic shape was significantly positively correlated with the thoracic translation distance and the bilateral ratio of the thoracic and iliocostalis muscles. In addition, the bilateral ratio of the thoracic iliocostalis muscles was significantly negatively correlated with the bilateral ratios of the lower thoracic shape and lumbar iliocostalis muscles. [Conclusion] Our findings showed that the asymmetry of the lower thoracic shape is associated with left lateral deviation of the thorax at rest and thoracic translation distance. In addition, the thoracic and lumbar iliocostalis muscle activity differed between the left and right translations.

Relationship between the thoracic asymmetry in standing position and the asymmetry of ankle moment in the frontal plane during gait.

[Purpose] We aimed to investigate the relationship of thoracic asymmetry in standing position with asymmetry of the internal ankle moment in the frontal plane during gait. [Participants and Methods] The following measurements were recorded in 22 healthy adult males using a 3D motion analyzer and force plates: thoracic lateral deviation, asymmetrical ratios of the upper and lower thoracic shape, internal ankle moment in the frontal plane, mediolateral deviations of the center of mass and center of pressure. [Results] In the standing position, the thorax was deviated to the left relative to the pelvis, and the upper and lower thoracic shapes were asymmetrical. During gait, significant lateralities were observed in the internal ankle moment in the frontal plane, mediolateral deviations of the center of mass and the center of pressure. Significant positive correlations were observed between the asymmetrical ratio of the lower thoracic shape and both the asymmetry of the internal ankle moment in the frontal plane and the mediolateral deviation of the center of pressure. [Conclusion] These results suggest that thoracic asymmetry is associated with mediolateral control of the ankle during gait.

Relationship between scapular elevation exercises with different alignments and activity of the trapezius and levator scapulae muscles.

[Purpose] This study aimed to examine whether scapular elevation exercises in sitting positions with different alignments lead to contractions of the trapezius and levator scapulae muscles. [Participants and Methods] The participants were 25 males, measured in four sitting positions with different alignments. Spine alignment was assessed by measuring the head protrusion, upper thoracic spine tilt, and pelvic tilt angles. Upper limb alignment was evaluated using the scapula tilt angle, scapula rotation angle, and distance between scapular spinous processes. Scapular elevation exercises were measured, and the thickness of the trapezius and levator scapulae muscles were measured in resting and elevated positions, with changes in muscle thickness. [Results] The trapezius muscle thickness was greater in the sitting position with less thoracic spine tilt and scapula tilt angles. Conversely, the levator scapulae muscle thickness was greater in the sitting position with more thoracic spine tilt and scapula tilt angles. [Conclusion] Scapular elevation exercises induce separate contractions of the trapezius and levator scapulae muscles by modifying the alignment of the spine and upper limbs.

Neonatal maternal separation causes decreased numbers of sertoli cell, spermatogenic cells, and sperm in mice.

Neonatal maternal separation is an experimental model used to evaluate the effects of toxic stress in neonates, or early life stress. Although various physiological and psychological stresses during childhood have been reported, the effects of neonatal maternal separation on the male reproductive system remain unclear. Therefore, the present study evaluated the effects of neonatal maternal separation on the male reproductive system. In neonatal male ICR mice, maternal separation was performed for 0.5, 1, 2, and 4 hours/day, from postnatal day 1 to 10. At 10 weeks of age, the neonatal maternal separation mice exhibited decreases in both testicular weight and epididymal sperm number, along with various testicular morphological changes involving germ cells, Sertoli cells, and interstitial cells. Notably, neonatal maternal separation mice showed decreased numbers of Sertoli cells. Animals subjected to 0.5-, 1-, and 2-h/day neonatal maternal separation exhibited decreases in serum levels of testosterone but not in those of gonadotropin (luteinizing hormone and follicle-stimulating hormone). Together, these data showed that neonatal maternal separation in male mice causes decreased Sertoli cell numbers following puberty, resulting in subsequent decreased spermatogenic activity.

Third primary branch of the posterior ramus of the spinal nerve at the thoracolumbar region: a cadaveric study.

PURPOSE: Spinal column procedures require an accurate understanding of neural pathways relative to the anatomic structure. Since Bogduk's report in 1982, it has been known that the human lumbar posterior ramus of the spinal nerve (PRSN) comprise not two but three primary branches at least in some lumbar segments. The purpose of the current study was to examine the existence of the three primary branches in the thoracic and lumbar segments. METHODS: In this study, we investigated the anatomy of the human PRSN in the thoracic and lumbar segments. Ventral dissection was performed in eight cadavers to determine the anatomy of the PRSN between T1 and L5. RESULTS: At the distal end of a given PRSN, the PRSN divided into three primary branches-medial, intermediate and lateral-in 196 out of 272 segments in the thoracic and lumbar regions in eight cadavers. The medial branch supplied the spinalis compartment, and reached the skin. The lateral branch supplied the iliocostalis muscle compartment, and reached skin. The intermediate branch supplied the longissimus muscle and the area between the medial and the lateral branch, which was a seemingly shorter branch. CONCLUSION: The triplication of the primary branch of the PRSN is considered not uncommon. The third branch should be recognized in the literature and in textbooks.

Recombinant human soluble thrombomodulin improved lipopolysaccharide/d-galactosamine-induced acute liver failure in mice.

The effect of recombinant human soluble thrombomodulin (TM-α) on acute liver failure (ALF) is unclear, and we elucidated the effect of TM-α in lipopolysaccharide (LPS)/d-galactosamine (GalN)-induced ALF in mice. Placebo (saline) or TM-α (100 mg/kg) was administered 1 h after LPS/GalN administration. Survival rates were evaluated for 24 h after LPS/GalN administration. Plasma and liver samples were evaluated 1, 3, and 7 h after LPS/GalN administration. Survival rates were significantly higher in the TM-α-treated group than in the placebo group. A significant augmentation of plasma high-mobility group box 1 protein (HMGB1) was observed 7 h after LPS/GalN administration. In the TM-α-treated mice, plasma HMGB1 was significantly lower than in the placebo group. A significant augmentation of hepatic nuclear factor (NF)-κB p65 was observed in the placebo-treated group, whereas a significant reduction, relative to placebo, was observed in the TM-α-treated group. Hepatic expression of tumor necrosis factor (TNF)-α and myeloperoxidase were significantly increased in the placebo group, and were similarly significantly attenuated in the TM-α-treated group. TM-α treatment also produced a significant attenuation of liver neutrophil accumulation after LPS/GalN administration. Thus, TM-α may become a useful treatment strategy for reducing the symptoms of ALF via the attenuation of LPS/GalN-induced HMGB1 levels.

Chymase inhibition attenuates lipopolysaccharide/ d-galactosamine-induced acute liver failure in hamsters.

BACKGROUND/AIMS: Chymase inhibition has been shown to attenuate matrix metalloproteinase (MMP)-9 and tumor necrosis factor (TNF)-α, both of which are associated with the pathogenesis of acute liver failure (ALF). This study investigated the effects of the chymase inhibitor TY-51469 on lipopolysaccharide (LPS)/D-galactosamine (GalN)-induced ALF in hamsters. METHODS: TY-51469 (10 or 30 mg/kg) or placebo was administered 1 h before the LPS (160 µg/kg)/GalN (400 mg/kg) injection. RESULTS: Hepatic chymase activity was significantly increased after the LPS/GalN injection, but the significant increase was dose-dependently and significantly attenuated by treatment with TY-51469. Significant increases in hepatic MMP-9 activity and TNF-α concentration were observed after the LPS/GalN injection, but these increases were also attenuated by treatment with TY-51469. Plasma aspartate aminotransferase and alanine aminotransferase activities were significantly increased after LPS/GalN injection in the placebo-treated group, but the increases were significantly attenuated in the TY-51469-treated group. The area of hepatic necrotic after LPS/GalN injection was significantly reduced by treatment with TY-51469. Treatment with TY-51469 resulted in significant reductions in the hepatic malondialdehyde concentration, mast cell numbers, and gene expressions of interleukin-1ß and myeloperoxidase. DISCUSSION: Chymase inhibition could be a useful strategy to attenuate LPS/GalN-induced ALF in hamsters.

Regulation of bcl-2 transcription by estrogen receptor-α and c-Jun in human endometrium.

The estrogen-estrogen receptor (ER) signaling pathway plays crucial physiologic roles in not only the control of reproduction, but also in the generation of cancer in the breast and uterus. While some ER target genes have been identified containing the estrogen-responsive element (ERE), others are activated eventually by ER via protein-protein interaction without binding to ERE. In a previous study, we identified that the proliferative phase-specific expression of the bcl-2 gene in glandular cells could be regulated by the binding of c-Jun to its motifs in the promoter. Results from our present study indicate that the menstrual cyclic expression of bcl-2 could be controlled by either direct binding of ERα to ERE in the c-Jun promoter or the interaction of ERα with c-Jun that binds to its motifs in the bcl-2 gene. Intriguingly, the transcriptionally active form of c-Jun phosphorylated at Ser63 was identified binding to its motifs in the bcl-2 gene in a menstrual cyclic non-specific manner. Our study revealed a novel mechanism that transcriptionally regulates the expression of bcl-2 in the normal human endometrium.

Effects of female bone marrow transplantation on male reproductive organs.

Graft-versus-host disease (GVHD), an adverse effect after bone marrow transplantation (BMT), may affect male reproductive function. It is hypothesized that a sex-mismatched BMT induces GVHD in male reproductive organs because female immune cells are not immunologically tolerant to specific antigens of the male organs. However, this hypothesis has not been experimentally verified using male (M) recipient animals following BMT from the female (F) donors. Therefore, the aim of the present study is to examine whether the female BMT to males (F→M group) induces some GVHD reactions in the testis and the other male reproductive organs. The results showed that no inflammation was found in recipients of the male BMT to males (M→M group), whereas significant inflammatory cell responses lasting for at least 4 months were induced in testis, epididymis, prostate and preputial gland in some mice of F→M group. The most severe lesion was found in the preputial gland, in which lymphocytic inflammation was accompanied by loss of glandular acini, thickening of the interstitum and increased cytokines such as TNF-α and IFN-γ. Western blot analyses revealed that sera from the F→M group reacted with various antigens of the male reproductive organs. These results indicate that transplanted female immune cells may recognize the male reproductive organs as immunologically foreign ones and induce chronic GVHD, which may affect male reproductive function.

Vascular anatomy of the lateral meniscus with special focus on the joint capsule.

Previous studies have reported that the lateral meniscus (LM) has two regions, the popliteal hiatus area (PH) with a scarce blood supply and the roots with an abundant one. However, the description of its vascular anatomy remains insufficient. We hypothesized that the difference in the width of the meniscus hilum (MH) affects the scarcity and abundance of blood supply to the LM. The MH is a concept proposed by us and is the only site of entrance or exit of blood vessels and nerves associated with the meniscus. The purpose of this study was to provide a structural explanation for the disparity of blood supply to the LM using the concept of MH. Sixteen knees were examined to investigate the blood supply to LM. In most areas, the femoral joint capsule (FJC) and tibial joint capsule (TJC) continued to the cranial and caudal edges of the LM, respectively. In the roots, the FJC and TJC covered the femoral and the outer-femoral surfaces. In contrast, the FJC in the PH did not attach to the cranial edge and only the TJC there did to the caudal edge of the LM. Histochemical examination showed that the blood vessels enter the LM via the MH. In the PH, the MH at the caudal edge was extremely narrow; and in the roots, the MH on the outer-femoral surfaces was wide. The results suggest that the difference in the width of the MH affected the scarcity and abundance of blood supply to the LM.

Significance of matrix metalloproteinase-9 inhibition by imidapril for prevention of abdominal aortic aneurysms in angiotensin II type 1 receptor-knockout mice.

To clarify the matrix metalloproteinase (MMP)-9 inhibitory effect of an angiotensin-converting enzyme (ACE) inhibitor in vivo, we evaluated the effect of an ACE inhibitor against elastase-induced abdominal aortic aneurysm (AAA) progression in mice. Molecular models showed that imidapril bound directly to the mouse MMP-9 active center. An active form of imidapril, imidaprilat, dose-dependently inhibited MMP-9 activity in the extract from elastase-induced AAA in wild-type mice. Imidapril (10 mg/kg per day) was administered to wild-type or angiotensin II type 1 (AT1) receptor knockout mice. Blood pressure was significantly lower in AT1 receptor-knockout mice than in wild-type mice, but imidapril did not affect blood pressure in AT1 receptor-knockout mice. The aortic diameter was significantly expanded after elastase application, but the expansion was significantly lower in AT1 receptor-knockout mice than in wild-type mice. In AT1 receptor-knockout mice, the aortic expansion was further attenuated by imidapril. MMP-9 activity in aorta was significantly augmented after elastase application. The MMP-9 activity was significantly lower in AT1 receptor-knockout mice than in wild-type mice, and it was further attenuated by imidapril. In conclusion, MMP-9 inhibition by imidapril might contribute to the attenuation of AAA progression in AT1 receptor-knockout mice.

Chymase inhibitor ameliorates hepatic steatosis and fibrosis on established non-alcoholic steatohepatitis in hamsters fed a methionine- and choline-deficient diet.

AIM: Chymase plays a role in the augmentation of angiotensin II formation, which is involved in liver fibrosis. The therapeutic effects of a chymase inhibitor, TY-51469, on established hepatic steatosis and fibrosis were investigated in a model of developed non-alcoholic steatohepatitis. METHODS: Hamsters were fed a normal diet or methionine- and choline-deficient (MCD) diet for 12 weeks. Then, treatment with TY-51469 (1 mg/kg per day) or placebo was initiated, and the treatment was continued concurrently with the MCD diet for an additional 12 weeks. RESULTS: At 12 weeks after initiating the MCD diet, marked hepatic steatosis and fibrosis were observed in MCD diet-fed hamsters. Malondialdehyde and gene expression levels of collagen I, collagen III, α-smooth muscle actin (α-SMA) and Rac-1 in liver extracts were also increased in the MCD-diet-fed hamsters at 12 weeks. At 24 weeks, hepatic steatosis and fibrosis were more prominent in the placebo-treated hamsters that were fed the MCD-diet for 24 weeks versus 12 weeks. Hamsters treated with TY-51469 for 12 weeks after being on a 12-week MCD diet had significant ameliorations in both hepatic steatosis and fibrosis, and there were no significant differences compared to normal diet-fed hamsters. There were significant augmentations in angiotensin II and malondialdehyde, and gene expressions of collagen I, collagen III, α-SMA and Rac-1 in the placebo-treated hamsters at 24 weeks; however, these levels were reduced to normal levels in the TY-51469-treated hamsters. CONCLUSION: TY-51469 not only prevented the progression of hepatic steatosis and fibrosis, but also ameliorated hepatic steatosis and fibrosis.

Pathophysiological consequences and treatment strategy of obstructive jaundice.

Obstructive jaundice (OJ) is a common problem in daily clinical practice. However, completely understanding the pathophysiological changes in OJ remains a challenge for planning current and future management. The effects of OJ are widespread, affecting the biliary tree, hepatic cells, liver function, and causing systemic complications. The lack of bile in the intestine, destruction of the intestinal mucosal barrier, and increased absorption of endotoxins can lead to endotoxemia, production of proinflammatory cytokines, and induce systemic inflammatory response syndrome, ultimately leading to multiple organ dysfunction syndrome. Proper management of OJ includes adequate water supply and electrolyte replacement, nutritional support, preventive antibiotics, pain relief, and itching relief. The surgical treatment of OJ depends on the cause, location, and severity of the obstruction. Biliary drainage, surgery, and endoscopic intervention are potential treatment options depending on the patient's condition. In addition to modern medical treatments, Traditional Chinese medicine may offer therapeutic benefits for OJ. A comprehensive search was conducted on PubMed for relevant articles published up to August 1970. This review discusses in detail the pathophysiological changes associated with OJ and presents effective strategies for managing the condition.

The mechanism of Yinchenhao decoction in treating obstructive-jaundice-induced liver injury based on Nrf2 signaling pathway.

BACKGROUND: Obstructive jaundice (OJ) is caused by bile excretion disorder after partial or complete bile duct obstruction. It may cause liver injury through various mechanisms. Traditional Chinese medicine (TCM) has a lot of advantages in treating OJ. The recovery of liver function can be accelerated by combining Chinese medicine treatment with existing clinical practice. Yinchenhao decoction (YCHD), a TCM formula, has been used to treat jaundice. Although much progress has been made in recent years in understanding the mechanism of YCHD in treating OJ-induced liver injury, it is still not clear. AIM: To investigate chemical components of YCHD that are effective in the treatment of OJ and predict the mechanism of YCHD. METHODS: The active components and putative targets of YCHD were predicted using a network pharmacology approach. Gene Ontology biological process and Kyoto Encyclopedia of Genes and Genomes path enrichment analysis were carried out by cluster profile. We predicted the biological processes, possible targets, and associated signaling pathways that YCHD may involve in the treatment of OJ. Thirty male Sprague-Dawley rats were randomly divided into three groups, each consisting of 10 rats: the sham group (Group S), the OJ model group (Group M), and the YCHD-treated group (Group Y). The sham group only received laparotomy. The OJ model was established by ligating the common bile duct twice in Groups M and Y. For 1 wk, rats in Group Y were given a gavage of YCHD (3.6 mL/kg) twice daily, whereas rats in Groups S and M were given the same amount of physiological saline after intragastric administration daily. After 7 d, all rats were killed, and the liver and blood samples were collected for histopathological and biochemical examinations. Total bilirubin (TBIL), direct bilirubin (DBIL), alanine aminotransferase (ALT), and aspartate transaminase (AST) levels in the blood samples were detected. The gene expression levels of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS), and the nucleus positive rate of NF-E2 related factor 2 (Nrf2) protein were measured. Western blot analyses were used to detect the protein and gene expression levels of Nrf2, Kelch-like ECH-associated protein 1, NAD(P)H quinone dehydrogenase 1 (NQO1), and glutathione-S-transferase (GST) in the liver tissues. One-way analysis of variance was used to evaluate the statistical differences using the statistical package for the social sciences 23.0 software. Intergroup comparisons were followed by the least significant difference test and Dunnett's test. RESULTS: The effects of YCHD on OJ involve biological processes such as DNA transcription factor binding, RNA polymerase II specific regulation, DNA binding transcriptional activator activity, and nuclear receptor activity. The protective effects of YCHD against OJ were closely related to 20 pathways, including the hepatitis-B, the mitogen-activated protein kinase, the phosphatidylinositol 3-kinase/protein kinase B, and tumor necrosis factor signaling pathways. YCHD alleviated the swelling and necrosis of hepatocytes. Following YCHD treatment, the serum levels of TBIL (176.39 ± 17.03 µmol/L vs 132.23 ± 13.88 µmol/L, P < 0.01), DBIL (141.41 ± 14.66 µmol/L vs 106.43 ± 10.88 µmol/L, P < 0.01), ALT (332.07 ± 34.34 U/L vs 269.97 ± 24.78 U/L, P < 0.05), and AST (411.44 ± 47.64 U/L vs 305.47 ± 29.36 U/L, P < 0.01) decreased. YCHD promoted the translocation of Nrf2 into the nucleus (12.78 ± 0.99 % vs 60.77 ± 1.90 %, P < 0.001). After YCHD treatment, we found a decrease in iNOS (0.30 ± 0.02 vs 0.20 ± 0.02, P < 0.001) and an increase in eNOS (0.18 ± 0.02 vs 0.32 ± 0.02, P < 0.001). Meanwhile, in OJ rats, YCHD increased the expressions of Nrf2 (0.57 ± 0.03 vs 1.18 ± 0.10, P < 0.001), NQO1 (0.13 ± 0.09 vs 1.19 ± 0.07, P < 0.001), and GST (0.12 ± 0.02 vs 0.50 ± 0.05, P < 0.001), implying that the potential mechanism of YCHD against OJ-induced liver injury was the upregulation of the Nrf2 signaling pathway. CONCLUSION: OJ-induced liver injury is associated with the Nrf2 signaling pathway. YCHD can reduce liver injury and oxidative damage by upregulating the Nrf2 pathway.

Neonatal corticosterone administration increases p27-positive Sertoli cell number and decreases Sertoli cell number in the testes of mice at prepuberty.

Cortisol and corticosterone (CORT) are steroid, antistress hormones and one of the glucocorticoids in humans and animals, respectively. This study evaluated the effects of CORT administration on the male reproductive system in early life stages. CORT was subcutaneously injected at 0.36 (low-), 3.6 (middle-), and 36 (high-dosed) mg/kg body weight from postnatal day (PND) 1 to 10 in ICR mice. We observed a dose-dependent increase in serum CORT levels on PND 10, and serum testosterone levels were significantly increased only in high-dosed-CORT mice. Triiodothyronine levels were significantly higher in the low-dosed mice but lower in the middle- and high-dosed mice. However, testicular weights did not change significantly among the mice. Sertoli cell numbers were significantly reduced in low- and middle-dosed mice, whereas p27-positive Sertoli cell numbers increased in low- and middle-dosed mice. On PND 16, significant increases in testicular and relative testicular weights were observed in all-dosed-CORT mice. On PND 70, a significant decrease in testicular weight, Sertoli cell number, and spermatozoa count was observed. These results revealed that increased serum CORT levels in early life stages could induce p27 expression in Sertoli cells and terminate Sertoli cell proliferation, leading to decreased Sertoli cell number in mouse testes.

Characteristic expression of CGRP and osteogenic and vasculogenic markers in the proximal and distal regions of the rib during male mouse development.

BACKGROUND: Neuropeptide calcitonin gene-related peptide (CGRP) is a neurotransmitter related to vasculogenesis and osteogenesis during bone formation and organ development. From the foetal period to the postnatal period, the thorax, which is necessary for lung respiration, forms. The thorax exhibits the same cartilage ossification as the bones of the extremities, but a specific system within the thorax exists as costal cartilage after birth. The relationship among CGRP, osteogenesis and vasculogenic markers in the two rib locations during thorax formation is not fully understood. MATERIALS AND METHODS: In our study, male mice were used to provide ribs under different development conditions on various embryonic days (E12. 5, E14.5, and E17.5) and postnatal days (P1 and P5). The mRNA expression levels of CGRP, vascular endothelial growth factor (VEGF-A), type 1 collagen (Col1a-1), type 2 collagen (Col2a1), neuropeptide Y (NPY), osteocalcin (OCN) and osteopontin (OPN) were analysed by qRT-PCR. We also analysed the mRNA expression of CGRP, VEGF-A and OPN by in situ hybridization. Multivariate modelling with principal component analysis (PCA) was performed to estimate the interactions among the quantitative real-time RT-PCR data. RESULTS: The mRNA expression levels of CGRP, VEGF-A, Col2a, Col1a-1, OCN, and NPY in the male mouse rib gradually increased during development. An antisense probe for CGRP mRNA was strongly detected in the central region of the mouse rib at E12.5 and the hypertrophic and ossification zones at E17.5 by in situ hybridization. VEGF-A was also located in the same region as CGRP at E12.5 and E17.5. OPN was strongly detected at the rib formation stage from E14.5 to E17.5. The expression of CGRP also differed between the proximal and distal regions of the rib at E17.5. As demonstrated by in situ hybridization, CGRP continuously participates in cartilage formation in the distal regions of the rib after birth. The PCA revealed that the mRNA expression of CGRP was related to that of Col1a-1 and VEGF-A during rib formation. CONCLUSION: This study shows that CGRP is involved in vascular and bone formation during rib development and may also be involved in cartilage formation after birth. The findings suggest that CGRP may temporarily participate in bone formation and continuously participate in cartilage formation in the rib, which may also be related to the formation of the anterior thoracic wall after birth.

Analysis of the mylohyoid nerve in elderly Japanese cadavers for dental implant surgery.

OBJECTIVES: Injury to the mandibular nerve (MN) branches may cause pain and irregular occlusal movement during mastication after mandibular dental treatments. Growing evidence indicates that the calcitonin gene-related peptide (CGRP) plays a key role in the development of peripheral sensitization and the associated enhanced pain, suggesting it may be a sign to ensure a safe and reliable dental implant treatment. Our focus was on the distribution of the MN branches and their communication with the lingual nerve (LN), the localized expression of CGRP, and the identification of a pain area related to the mylohyoid muscle (MM) fascia in the mandibular floor. MATERIAL AND METHODS: In this study, MM samples from 440 sides of 303 human cadavers aged 61-103 years were examined microscopically and immunohistochemically. These data were further evaluated by the use of principal component analysis. RESULTS: A complex but weak attachment site was identified for the fascia of the MM. CGRP expression was mainly located in small vessels and was scattered throughout the whole fascia of the MM. Communication between the MN and LN was found in 62.5% (275/440) of the samples. The results from the principal component analysis showed that the positive contributions were from the descending branch in the premolar region (correlation coefficient value R = 0.665), the ascending branch in the molar region (R = 0.709) and the intermediate branch of the digastric branch (R = 0.720) in component 1. In the fascia off the MM, strongly labeled CGRP-positive cells were also found around the blood vessels and the nerve. CONCLUSIONS: The findings reported in this study indicate that there is a risk of damage when pulling the fascia off the MM at the border of the molar and premolar regions during dental implant surgery.

Comparison of CGRP distributions in the maxillary sinus and trigeminal ganglion between elderly dentulous and edentulous humans.

Thickening of the Schneiderian membrane (SM, mucosa of the maxillary sinus) appears in the paranasal sinus. Information on SM thickening is available for patients receiving sinus lift treatments, which is a risk factor for SM excretory dysfunction. However, more information is needed on the structure of the SM and the relationship between the maxilla sinus and palatine with the alveolar bone and the SM for dental implant treatment in the human maxilla. One hundred twenty-six sides of the maxilla from 71 cadavers were subjected to cone-beam CT (CBCT) analysis and macroscopic and immunohistochemical observations in this study. A thickened SM was mainly observed in the middle region of the basal layer of the maxillary sinus (MS). Strong calcitonin gene-related peptide (CGRP)-positive reactions were observed in the alveolar bone, oral mucosa, mucosa of the maxillary sinus, and trigeminal ganglion (TG) cells in dentulous samples compared with edentulous samples. TG cells play important roles in delivering CGRP through axons to the mucosal gland and in regulating the maxilla-related thickening of the SM. These data could help determine CGRP functions in the mucosal gland and bone formation between dentulous and edentulous samples and indicate that CGRP may pass from the TG to the maxillary sinus glands.