Tracheobronchial foreign body aspiration can cause serious health problems in individuals of all age groups and requires immediate diagnosis and intervention. Patients with psychiatric disorders can aspirate foreign objects that may pose a danger to their health. Herein, we report on a case of curtain clip aspiration in a 44-year-old man with mental retardation. Chest radiography and bronchoscopy showed a sharp, pointed, S-shaped curtain clip inversely lodged in his left main bronchus. The curtain clip was removed successfully using flexible bronchoscopy, and no complications were noted.

In order to monitor the developmental features of embryos, larvae, and juveniles of Oryzias latipes (Temminck and Schlegel), Oryzias latipes was caught in river of Shinduck-dong, Yeosu-si, Jeollanam-do, on May 2011, and experiments were carried out in Ichthyology laboratory at Chonnam National University. The blastodisc step was the first level for natural spawning. The optic vesicle, Kupffer's vesicle, myotome began to appear 75 hours 57 minutes later. After blastodisc development, the pectoral fins were made at 143 hours 37 minutes and the tail was separated started at the same time. Hatching was observed at 167 hours 27 minutes after blastodisc. The total length of the hatched larvae was 4.95~5.10 mm (mean, 5.01 mm), the mouth and anus were opened. Larvae used yolk completely after 3 days after hatching. The total length larvae was 5.45~5.56 mm (mean, 5.52 mm) after 8 days after hatching, and appeared the stems for tail. The stems pectoral, anal fin were showed after 14 days and the stems dorsal, ventral fin were appeared after 19 days. For 35 days after hatching, the total length of larvae 13.95~15.30 mm (mean, 14.64 mm), and at this time, fins and body were transferred like the adult Oryzias latipes.

Cholesterol embolization syndrome (CES) is a multiple systemic disease caused by the embolization of cholesterol crystals from an atherosclerotic plaque of a proximal large-caliber artery, which results in the occlusion of distal small to mediumsized arteries. CES is characterized by development of a multitude of small emboli over time, and should be distinguished from arterial thromboembolism, which occurs through the obstruction of medium-sized to large arteries by one or a few large emboli. We report on a case of CES initially presenting as acute limb ischemia following an intervention for iliac artery occlusion.

The isolated single coronary artery is a rare congenital anomaly, in which both coronary arteries arise from a solitary ostium. Diagnosis of coronary anomalies and identification of the exact anatomy of coronary arteries has significant clinical importance, hence, myocardial ischemia or sudden cardiac death is usually related to its course of anomalous coronary artery. Most patients with a single coronary artery are asymptomatic and have normal electrocardiogram and negative stress tests. However, if the patient has other structural abnormalities, for example, ventricular hypertrophy, the exam is determined. This report describes a case of single coronary artery, where the right coronary artery originated from the distal left circumflex artery in a patient with hypertrophic ardiomyopathy.

We investigated the androgenic effects of 17α-methyltestosterone (MT) on gonadal sex reversal in juvenile red spotted grouper Epinephelus akaara. The fish were immersed in 17α-MT at 1 and 5 mg/L. Treatment method of 17α-MT was once weekly for 4 and 8 weeks. Fish were sampled at 12 months after end of the treatment period in order to histological analysis. At the initiation of an experiment (70 day after hatching), juvenile red spotted grouper have the paired primordial gonads with somatic cells bellow kidney in the posterior portion of the body cavity. Formation of ovarian cavity indicates that the ovarian differentiation beginning at 70 DAH in red spotted grouper. At 12 months after end of the treatment period, control group, 17α-MT 1 mg/L treatment group for 4 and 8 weeks, and 17α-MT 5 mg/L treatment group for 4 weeks were all female. However, sex-changed males without ovarian cavity were observed in the 17α-MT 5 mg/L treatment group for 8 weeks. In grouper, we firstly reported that the red spotted grouper be able to induce the primary males by hormone treatment prior to gonadal sex differentiation.

Estrogen is an important regulator of reproduction in both male and female. The two forms of estrogen receptor (ER) are known, ERα and ERβ. To understand the role of ERα in the testis, we investigated the expression of ERα in the mouse Leydig cells during postnatal development and the effects of estrogen on steroidogenesis and proliferation in progenitor Leydig cells (PLCs). In the testis, ERα mRNA and protein levels were markedly increased from postnatal day (PND) 1 to 14 and decreased thereafter until PND 56. During postnatal development ERα immunoreactivity was strong in the nucleus of Leydig cells at PND 14 when PLCs were abundant in the interstitium and low in the mature adult Leydig cells (ALCs). In fetal Leydig cells (FLCs), ERα immunoreactivity was negligible at birth and became increased at PND 14. This suggests an important role of ERα in Leydig cells during neonatal period. In isolated PLCs, 17β-estradiol (E2) and ERα-selective agonist, PPT suppressed the hCG-induced progesterone production and steroidogenic pathway genes expression. The hCG-induced PLCs proliferation was significantly inhibited by E2 and PPT. In conclusion, estrogen - ERα signaling may negatively regulate functional differentiation and proliferation of PLCs.

Reactive oxygen species (ROS) are produced in organisms as the natural products of oxidative metabolism by environmental stress and pathogen invasion. ROS, such as superoxide anion and hydrogen peroxide, can be toxic to cells and tissues to cause oxidative stress. Recent study revealed that olive flounder (Paralichthys olivaceus) superoxide dismutase (SOD) has been identified as a partial gene and strongly induced to benzoin[a]pyrene and it was deduced indicator of aquatic oxidative stress responses, but its transcriptional response against viral infection has not been investigated. In the present study, spatial and temporal expression profile was analyzed to investigate the function of Of-SOD in the anti-viral response. Of-SOD transcripts were ubiquitously detected in diverse tissues with variable levels using a real-time PCR. The expression of Of-SOD was significantly higher in the muscle, liver and brain, but extremely low in the stomach and spleen. Following VHSV challenge, the expression of Of-SOD increased within 3 hours and subsequently decreased to the original level at 2 days post-challenge in kidney. Although expression pattern and induction time are slight differences depending on the tissue, the transcript of Of-SOD was consistently increased in acute infection response, but expression is low in the chronic response. Collectively, Of-SOD expressions were inducible after VHSV infection and they were probably involved in the immune response against viral challenge. These results suggest that SODs may play important roles in the immune defense system of P. olivaceus and perhaps contribute to the protective effects against oxidative stress in this flounder.

Molecular markers are useful for selecting to include superior character genetic like as strong immune system and rapid growth in fish. The marker is also very important part of breeding technology in Olive flounder (Paralichthys olivaceus). Single nucleotide polymorphisms (SNPs) marker is already in use widely for genomic research and breeding. But this SNPs marker hardly has been validated for screening functional genes in Olive flounder. We study identify single nucleotide polymorphisms (SNPs) on Expressed sequence tag (EST) database, develop usable SNP marker and apply to wild sample and cultured of olive flounder. As a result, Out of total 4.327 ESTs, 693contigs and 514 SNP from total contigs were detected while these substitutions include 297 transitions and 217 transversions. 144 developed markers were applied in 16 samples (wild 8, culture 8), Out of total marker, only 32 markers had detected polymorphic in sample. Polymorphism of 32 markers was observed in the variety genes region involved in immunity and protein synthesis. And the 32 marker were identified 21 transitions, 11 transversions, and indel was not detected in polymorphic SNPs. The analysis on heterozygosity by sample showed 0.34 in wild sample and 0.29 in cultured sample. In conclusion, we was identified SNP and Polymorphism by designed new marker, it supports that development marker is suitable for SNP detection and diversity analysis in Olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

Multipotent perivascular stem cells (PVCs) have gained much attention as an alternative source for cell based regenerative medicine in recent years. Due to their rarity in human tissues, developing methods to efficiently isolate and expand PVCs from various fetal and adult tissues is necessary to obtain a clinically relevant number of cells that maintain progenitor potency. Here, we report on a non-enzymatic isolation (NE) method of PVCs from human umbilical cords (HUCs) and compare its efficiency with the conventional collagenase treatment method (CT) in terms of proliferation and immunophenotypes. The cells isolated by NE displyed acceptable surface marker profile of PVCs and showed multilineage (osteogenic, chondrogenic, and adipogenic) differentiation potential. While both methods provided similar levels or patterns of proliferation and immunophenotypes, PVCs by NE retained a higher level of CD146(+) frequency compared to that of CT over passage. Furthermore, we have investigated potentials of various exogenous factors to promote proliferation of HUCPVCs in vitro. Among these factors, supplementation of basic fibroblast growth factor (bFGF) provided the optimal condition to significantly enhance the proliferation rate of HUCPVC and increased a proportion of stage-specific antigen-4 (SSEA-4) positive subset. Collectively, our study suggests that NE method with bFGF supplementation offers an alternative way to obtain sufficient numbers of HUCPVCs with higher number of primitive SSEA-4(+) subpopulation that are applicable in therapeutic doses for regenerative medicine.

Melatonin has several known physiological functions, the main one being synchronization of daily and seasonal rhythms. In addition, melatonin has been reported to influence reproduction and behavioral rhythms with varying results depending on the species. To date, it remains unknown how this rhythm in locomotor activity is controlled endogenously, although there must be coordination of chemical and molecular drivers. However, the species is poorly characterized at molecular level with little sequence information available in public databases. The aim of study was to clarify involvement of endogenous melatonin rhythms and locomotor activity in day-night activity of the eel, Anguilla japonica which is an economically important but endangered species. The levels during daytime (zeitgeber time; ZT 6) were significantly (P<0.05) lower than those during nighttime (ZT 18). A similar pattern was persisted under DD conditions, whereas it disappeared under LL conditions and ocular melatonin levels remained low. Therefore, it is likely that ocular melatonin levels of the nocturnal eel reared under LD and DD conditions fluctuate in a daily/circadian manner and night-related physiological processes are dependent on eel locomotor activities which is a nocturnal species. We found that similar number of genes were differentially expressed between day (ZT6) and nighttime (ZT18), suggesting that during the nighttime also important in differential gene expression with daytime. This work also provides essential information for further studies investigating the molecular basis of daily/circadian system in this species.

As a preliminary investigation into the effect of environmental factors control for gonadal development, we examined the involvement of photoperiod and water temperature in ovarian development of Epinephelus. akaara. For the induction of sexual maturation, E. akaara reared in recirculating aquaculture system (RAS). During November 2013, the photoperiod and water temperature was adjusted to 12L:12D and 18℃, respectively. In the photo-thermal treatment group, every 3 weeks daylight was increased as follows a 13L:11D and 14L:10D, and control group was maintained under natural condition. After 9 weeks, water temperature was increased 23℃ in photo-thermal treatment group. The sampled fish every 3 weeks revealed increase in gonadosomatic index (GSI; 5.18±1.38), oocyte diameter and vitellogenic oocytes (423.9±36.1 ㎛) were observed in gonads 12 weeks under photo-thermal treatment group. However, ovarian development was maintained immature stage in control group. In this environmental factors manipulation trial, seventy one of the 95 females (578.4 ± 25.4 g in mean body weight, 31.0 ± 0.5 cm mean total length) treated with HCG injection (doses 500 IU/kg BW) were induced ovulation by artificial stripping. The total volume of ovulated eggs were 3,470 ml and the total volume of fertilized eggs was 3,295 ml. The fertilization rate and hatching rate were 95% and 98%, respectively. These results suggest that the photoperiod as well as water temperature are major environmental factors in triggering the gonadal development of E. akaara.

Aromatase is an enzyme that converts testosterone to estrogen. This enzyme, present in the sperm as well as various tissue and cells, has been considered to be related to the fertility of human and mouse sperm. Therefore, we examined effect of aromatase inhibitor on viability and fertility of sperm, and quantity of aromatase in sperm groups with different density in pig. To analyze the effect of aromatase on sperm viability, we treated aromatase inhibitor to the sperm with different concentrations (0, 10, 20, 50, 100, 200, 500 μM) at different time (0.5, 1, 2, 4, 8 hours). After the treatment, the sperm viability was calculated by hypo-osmotic swelling test. We selected 0, 50, 100 μM concentration during 0.5 hour as inhibitor treatment condition before in vitro fertilization. Next, we examined fertility and quantified aromatase protein in sperms with different density. In the first experiment, viability of sperm was decreased following the increasement of inhibitor concentration. The aromatase inhibited sperm showed lower penetration rate and cleavage rate than those of non-treated sperm. Concentration of 50 μM inhibitor had no significant effect on the sperm viability, but it significantly reduced sperm fertility. Second, sperms with low density showed higher penetration rate, but no significant difference between sperms with high density. In conclusion, aromatase is responsible for viability and fertility of porcine sperm similar to mouse and human, however, density of sperm has no correlation with quantity of aromatase protein.

Sirt1 belongs to class III histone/protein deacetylase. Sirt1 KO mice have spermatogenic defects. In this study, expression of Sirt1 and acetylated histone 3k9 (H3k9ac) was examined in developing mouse testes. Among two splicing variants of sirt1 mRNA long form was dominant in developing testis. Testicular sirt1 mRNA levels were low at birth, increased until 14 days post partum (pp) and remained constant thereafter. Sirt1 immunoreactivity was weak to negligible in gonocytes, moderate in spermatogonia, absent in preleptotene spermatocytes, moderate in zygotene spermatocytes, strong in pachytene spermatocytes, and weak in diplotene spermatocytes. Round and elongating spermatids were negative for Sirt1. Acetylated histone 3k9 (H3k9ac) immunoreactivity was moderate in spermatogonia and weak to negligible in preleptotene to pachytene spermatocytes but strong in round and elongating spermatids. In Sertoli cells, nuclear Sirt1 immunoreactivity was absent at birth, increased at 14 days pp and markedly increased at 28 days pp onwards. In immature Sertoli cells culture, FSH and testosterone increased sirt1 mRNA, suggesting that Sirt1 participates in protein deacetylation events during the differentiation of Sertoli cells by gonadotropin. In the Leydig cells, nuclear Sirt1 immunoreactivity was weak until 2 weeks pp and decreased in 4 weeks pp onward. suggesting the protein deacetylation by Sirt1 in Leydig cell precursor/progenitor cells. Mutually exclusive expression between Sirt1 and H3k9ac in pachytene spermatocytes in testis suggests that deacetylation of H3K9ac by Sirt1 participates in the gene silencing and/or chromosome behavior in pachytene sspermatocytes.

Water channel proteins, aquaporins (AQPs) contribute to transepithelial water movement in many tissues. To date, 13 mammalian AQPs have been identified. Of these, AQP5 plays an important role in the fluid homeostasis and cell volume control in epithelial cells. In an effort to understand the role of AQP5 in testis, we investigated the expression of AQP5 in developing mouse testis, its regulation by estrogen and LH, and the change of steroidogenesis by AQP5 knockdown. Testes and Leydig cells were isolated from male mice at postnatal day (PND) 1, 7, 14, 28, and 56 and estrogen receptor alpha knockout (ERαKO) mice. In mouse testis, AQP5 immunoreactivity was negligible by PND 14. From PND 28 onward, AQP5 immunoreactivity was found in Leydig cells. In ERαKO mouse Leydig cells, AQP5 mRNA level was significantly lower than wild type. In primary adult Leydig cell culture, the expression of AQP5 mRNA was increased by 17β-estradiol (E2) and human chorionic gonadotropin (hCG), but was not changed in ERαKO Leydig cells. Moreover, the expression of AQP5 mRNA was increased by E2 and ERα-selective agonist PPT, but was not changed by ERβ-selective agonist DPN in primary Leydig cells and mLTC-1. In silico analysis and chromatin immunoprecipitation (ChIP) assay revealed that there are putative estrogen response elements (EREs) and cAMP response elements (CRE) in AQP5 promoter region. Testosterone secretion and steroidogenic pathway genes (StAR, Cyp11a1, Cyp17a1, and 3β-HSD6) expression were decreased by AQP5 siRNA in primary Leydig cells. In conclusion, AQP5 expression was coupled with functional differentiation of adult Leydig cells. AQP5 may play an important role in the fluid homeostasis and cell volume control during development of adult Leydig cells. The expression of AQP5 in Leydig cells could be regulated by ERα and LH signaling and AQP5 may be involved in steroidogenesis.

Amnionless (AMN) is a plasma membrane protein that binds to cubilin and megalin in various epithelia and mediates endocytosis of extracellular ligands. This function has been studied in the kidney where it plays a key role in vitamin B12 and vitamin D homeostasis. Present study aimed to elucidate developmental pattern of expression of AMN during the peri-implantation period in mouse embryos. In an effort to understand functional role of AMN in the histiotropic nutrition in blastocyst, endocytotic function of AMN for apoplipoprotein was examined in blastocyst. Eight-week-old female mice were superovulated by intraperitoneal injection of 5 IU PMSG and 5 IU hCG 48h later. To obtain embryos, females were mated with males. Mouse embryos were collected at 12, 48, 56, 65, 72 and 96 h post-hCG by flushing oviducts and uterus, and we also obtained gestation day 6.5, 7.5 and 8.5 embryos in uterus. All samples were subjected to quantitative RT-PCR, whole-mount immunofluorescence and immunohistochemistry analysis. To analyze endocytotic function of AMN, we examined uptake experiment of FICT labeled apolipoprotein A-I (ApoA-I-FITC) following functional blocking of AMN in blastocysts. AMN and cubilin mRNA was expressed in all developmental stages of mouse embryos. Megalin was the first detected at morula stage. AMN protein was expressed in trophectoderm (TE) and inner cell mass (ICM). AMN and cubilin were expressed in visceral endoderm of GD 6.5 and 7.5 embryos and visceral yolk sec of GD 8.5 embryos. In normal IgG treated embryos, ApoA-I-FITC was detected in intracellular vesicles of TE and ICM. However, in the presence of anti-AMN antibody, ApoA-I-FITC was weakly detected in apical surface of plasma membrane of TE. To date, AMN has been believed to be expressed in visceral endoderm of post implantation embryos. Our results demonstrated that AMN is the important molecular partner of cubilin and megalin in the preimplantation embryos and that AMN mediates endocytosis of apoplipoprotein, which may play a crucial role in embryonic development and normal growth via supporting histiotropic nutrition during peri-implantation period.

Coxsackievirus and adenovirus receptor (CAR) is a member of the Ig-type superfamily of cell adhesion molecules. In polarized epithelial cells CAR is expressed at the tight junction. The mouse CAR gene is composed of at least eight exons, and CAR splice variants that differ at the end of the cytoplasmic tail have been identified in a number of tissues. The present study aimed to examine the expression of (CAR), a TJ protein sealing the muticellular contact point during preimplantation embryos and role of CAR in the formation and integrity of the blastocoel. Eight-week-old female mice were superovulated by intraperitoneal injection of 5 IU PMSG and 5 IU hCG 48h later. Mouse embryos were collected at 12, 48, 56, 65, 72 and 96 h post-hCG in oviducts and uterus, and we also obtained gestation day 5 and 6.5 embryos in uterus. All samples were subjected to RT-PCR, immunofluorescence and immunohistochemistry analysis. To analyze epithelial permeability of CAR, we examined permeability of FITC-labeled dextran (MW 40 kDa) following functional blocking of CAR in blastocysts. Long isoform of CAR mRNA was expressed from throughout the preimplantation development and markedly increased at morulae stage onward. Small amount of short isoform CAR mRNA was expressed at blastocyst stage. On Western blot, 64 kDa protein was detected together with 43 kDa protein corresponding to short and long forms CAR, respectively in blastocysts. CAR immunoreactivity was found in cell contacts between blastomeres from 4-cell stage onward. Under Ca2+ switching condition blocking antibodies for CAR increased the permeability of blastocysts to FITC-dextran, a permeability tracer. At 5 dpc, trophoblasts of the implanting embryos were immunoreactive with anti-CAR IgG. At 6.5 dpc, the egg cylinder stage in mouse, the visceral and parietal endoderm were immunoreactive with anti-CAR IgG. Our results suggest that alternative splicing of CAR transcript is highly dependent on the development of expanding blastocyst. CAR may play a crucial role as a barrier to adenovirus infection and adhesion molecule for epithelial permeability during peri-implantation period.

We investigated the change mRNA expression of GtHs subunits (FSHβ, LHβ) in the pituitary, androgen receptor (AR), estrogen receptor (ERα) in gonad and histological observation of gonads in longthooth grouper Epinephelus bruneus by treatment Femara, an aromatase inhibitor (AI). Longtooth grouper (body weight 408±43.1 g; one year) cultured in Future Aquaculture Research Center, NFRDI were used in the experiments. The experiment was conducted for 12 weeks from 21 August 2013. Fish received intramuscular injection of AI at 5 mg/g BW dose in three times every 3 weeks. Fish were sampled pituitary and gonads at 3, 6, 12 weeks post-injection (n=50). The mRNA levels of FSH-β, LH-β in pituitary and AR, ERα mRNA in gonad were evaluated using qRT-PCR and qPCR. The histological change of gonads observed on light microscope. The gonads of control group contained most perinucleolus oocyte. At 3 to 6 weeks post-injection, the gonads of AI-treated group contained a few degenerated oocytes, spermatogonia and spermatocytes. At 12 weeks post-injection, gonads contained spermatids undergoing spermatogenesis. From 6 to 12 weeks post-injection, the expression level of GtHs subunits mRNA in pituitary was significantly higher than control group. The expression level of AR mRNA in gonad was higher than control group from 3 to 12 weeks post-injection. The expression level of ERα mRNA in gonad was lower than control group from 6 to 12 weeks post-injection. These results suggest that immature longtooth grouper with AI treatment induced masculinization via change of GtH subunits in pituitary, AR and ERα mRNA in gonad.