• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.269-275
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• 1996

Biosynthesis and secretion of anterior pituitary hormones are under the control of specific hypothalamic stimulatory and inhibitory factors. Among them, Growth Hormone Releasing Hormone (GHRH) is the major stimulator of pituitary somatotrophs activating GH gene expression and secretion. Human GHRH is a polypeptide of 44 amino acids initially isolated from pancreatic tumors, and the gene for the hypothalamic form of GHRH is organized into 5 exons spanning over 10 kilobases (kb) on genomic DNA and encodes a messenger RNA of 700-750 nucleotides. Several neuropeptides classically associated with the hypothalamus have been found in the extrahypothalamic regions, suggesting the existence of novel sources, targets and functions. GHRH-like immunoreactivity has been found in several peripheral sites, including placenta, testis, and ovary, indicating that GHRH may also have regulatory roles in peripheral reproductive organs. Furthermore, higher molecular weight forms of the GHRH transcripts were identified from these organs (1.75 kb in testis; 1.75 and >3 kb in ovary). These tissue-specific expression of GHRH gene suggest the existence of unique regulatory mechanism of GHRH expression and function in these organs. In fact, placenta-specific and testis-specific promoters for GHRH transcripts which are located in about 10 kb upstream region of hypothalamic promoter were reported. The use of unique promoters in extrahypothalamic sites could be refered in a different control of GHRH gene and different functions of the translated products in these tissues. Somatotrophs and lactotrophs have been thought to be derived from a common bipotential progenitor, the somatolactotrophs, which give origins to either phenotypes. Although the precise mechanism responsible for the lactotroph differentiation in the anterior pituitary gland has not been yet clalified, there are several candidators for the generation of lactotrophs. In human, the presence of GHRH peptides with different size from authentic hypothalamic form in the normal anterior pituitary and several types of adenoma were demonstrated. Recently our group found the existence of immunoreactive GHRH and its transcript from the normal rat anterior pituitary (gonadotroph> somatotroph> lactotroph), and the GHRH treatment evoked the increased proliferation rate of anterior pituitary cells in vitro. The transgenic mouse models clearly shown that GHRH or NGF overexpression by anterior pituitary cells induced development of pituitary hyperplasia and adenomas particularly GH-oma and prolactinoma. Taken together, we hypothesize that the pituitary GHRH could serve not only as a modulator of hormone secretion but as a paracrine or autocrine regulator of anterior pituitary cell proliferation and differentiation. Interestingly enough, the expression of Pit-1 homeobox gene (the POU class transcription factor) was confined to somatotrophs, lactotrophs and somatolactotrophs in which GHRH receptors are expressed commonly. Concerning the mechanism of somatolactotroph and lactotroph differentiation in the anterior pituitary, we have focused following two possibilities; (1) changes in the relative levels or interactions of both hypothalamic and intrapituitary factors such as dopamine, VIP, somatostatin, NGF and GHRH; (2) alterations of GHRH-GHRH receptor signaling and Pit-1 activity may be the cause of lactotroph differentiation or pituitary hyperplasia and adenoma formation. Extensive further studies will be necessary to solve these complicated questions.

• The Journal of Korean Obstetrics and Gynecology
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• v.20 no.1
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• pp.99-113
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• 2007

Purpose : This study was designed to investigate the effects of Jeongkyeong-Tang(JKT) on the polycystic ovary(PCO) induced by estradiol valerate(EV) in rats. Methods : PCO was induced by single intramuscular injection with EV(4mg) in female rats. Normal group(n=8) were injected with sesame oil and orally administrated distilled water for sixty days. PCO control group(n=8) were injected with EV and orally administrated distilled water for sixty days. JKT treated group(n=8) were injected with EV and orally administrated JKT for sixty days. Then we measured weights of body, ovaries and adrenal glands, and measured content of serum estrogen. The histomorphometrical changes of ovaries were also evaluated. The expressions of nerve growth factor(NGF) were analyzed in the central nervous system, adrenal glands and ovaries by immunohistochemistry. Results : - The weights(mg) of ovaries in JKT treated group (69.7${\pm}$6.7) were significantly increased( p<0.001) compared with PCO control group(46.7${\pm}$12.2). - The numbers of secondary follicles in JKT treated group(4.00${\pm}$l.31) were significantly increased(p <0.05) compared with PCO control group(2.25${\pm}$1.39). - The numbers of mature follicles in JKT treated group(5.50${\pm}$1.51) were significantly increased(p<0.01) compared with PCO control group(2.88${\pm}$1.13). - The numbers of atretic follicles in JKT treated group(2.75${\pm}$l.16) were significantly decreased(p<0.001) compared with PCO control group(6.88${\pm}$2.03). - The numbers of corpora lutea in JKT treated group(4.13${\pm}$1.46) were significantly increased(p<0.01) compared with PCO control group(2.13${\pm}$1.46). - The contents(pg/ml) of serum estrogen in JKT treated group(115.18${\pm}$18.29) were significantly decreased(P<0.01) compared with PCO control group(153.06${\pm}$29.47). - The expressions of NGF-immunoreactive cells in the ovarian granulosa cells in JKT treated group were lesser observed than PCO control group. Conclusion : From the above results, we concluded that Jeongkyeong-Tang has inhibitory effect on the development of EV-induced polycystic ovary. And it's effect may be related with decreased NGF activities in the ovary.

• The Journal of Korean Obstetrics and Gynecology
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• v.25 no.2
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• pp.43-65
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• 2012

Objectives: This study was designed to investigate the effects of Hyeolbuchukeotang( HCT) on the Estradiol Valerate(EV) induced Polycystic Ovaries and the Implantation of rats. Methods: PCO was induced by single intramuscular(IM) injection with EV in female rats. Normal group(n=8) were injected with sesame oil and orally administrated distilled water for 60 days. Control group(n=8) were injected with EV and orally administrated distilled water for 60 days. HCT treated group(n=8) were injected with EV and orally administrated HCT for 60 days. At the end day of experiment, we measured weights of body, ovaries, adrenal glands, uterus and contents of serum LH, FSH, ADD. The histomorphometrical changes of ovaries were also evaluated. And we observed the NGF and CRF expression by immunohistochemistry. Also we observed the mating and count the number of implantation. Results: - The weights(mg) of ovaries in HCT treated group($58.9{\pm}6.5$) were significantly increased (p<0.01) compared with control group ($42.3{\pm}8.5$). - The number of mature follicles in HCT treated group($9.8{\pm}2.6$) was significantly increased (p<0.01) compared with control group($6.1{\pm}2.1$). - The number of cystic follicles in HCT treated group($1.6{\pm}1.2$) was significantly decreased (p<0.01) compared with control group($3.8{\pm}1.5$). - The value of serum FSH(mIU/ml) in HCT treated group($3.870{\pm}2.151$) was significantly decreased(p<0.05) compared with control group($10.476{\pm}6.294$). - The expression of NGF-immunoreactive cells in the ovarian cells in HCT treated group was lesser observed than control group. - The number of implantation in HCT treated group($7.4{\pm}1.8$) was significantly increased (p<0.05) compared with control group($3.1{\pm}4.4$). Conclusions: Hyeolbuchukeo-tang(HCT) is effect on polycystic ovaries and the implantation of rats by EV-induced.

• Journal of Veterinary Clinics
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• v.32 no.6
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• pp.544-547
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• 2015

An 11-year-old, intact male Pekingese was brought to the Veterinary Teaching Hospital of Kangwon National University with a 10-day history of seizures. Fifteen days before coming to Kangwon National University, the dog had visited a local animal hospital for lameness, and non-steroidal anti-inflammatory drugs were prescribed to treat this symptom. However, 10 days before coming to our hospital, the dog experienced generalized seizures. Two days before his arrival, generalized ataxia and mental dullness also occurred. Our examinations revealed no remarkable findings on a routine blood test or X-ray. However, the neurological examinations confirmed mental dullness, generalized ataxia, and a lack of menace response and pupillary light reflexes. Nine hours later, dyspnea occurred, and 12 hours after that, the patient was euthanized per the client's request. A necropsy of transverse sections confirmed the presence of a prominent midline shift due to extended tumor growth. On histopathological analyses, pseudopalisading necrosis of the glial cells and microvascular proliferation were observed. In immunohistochemical analysis, glial fibrillary acidic protein, proliferating cell nuclear antigens, and ionized calcium binding adaptor molecule 1 immunoreactive cells were observed in the tumor area. Based on the results, the tumor was confirmed to be a glioblastoma. Primary intracranial tumors are rare in the veterinary field. This case report describes the clinical and histopathological findings of glioblastoma in a Pekingese.

• The Journal of Korean Obstetrics and Gynecology
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• v.21 no.3
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• pp.60-74
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• 2008

Purpose: This study was designed to investigate the effects of Gwibi-Tang(GBT) on the polycystic ovary(PCO) induced by estradiol valerate(EV) in rats. Methods: PCO was induced by single intramuscular injection with EV(4mg) in female rats. Normal group(n=8) were injected with sesame oil and orally administrated distilled water for eight weeks. PCO control group(n=8) were injected with EV and orally administrated distilled water for eight weeks. GBT treated group(n=8) were injected with EV and orally administrated GBT for eight weeks. Then we measured weight of body, ovaries, adrenal glands, and uterus of rats. The histopathology changes of ovaries were also evaluated. The expression of nerve growth factor(NGF) was analyzed in the central nervous system, adrenal glands and ovaries by immunohistochemistry. And also CRF expression in median eminance of Rats were analyzed. Results: 1. The weight(g) of rats in GBT treated group($275{\pm}14$) was significantly increased(p<0.01) compared with control group($253{\pm}8$), 2. The weight(mg) of ovaries in GBT treated group($75.8{\pm}16.7$) was significantly increased(p<0.001) compared with control group($37.4{\pm}6.7$). 3. The number of mature follicles in GBT treated group($3.6{\pm}1.2$) was significantly increased(p<0.01) compared with control group($1.5{\pm}1.5$. 4. The number of atretic follicles in GBT treated group($8.0{\pm}3.1$) was significantly decreased(p<0.01) compared with control group($18.6{\pm}6.0$). 5. The number of cystic follicles in GBT treated group($0.5{\pm}0.5$) was significantly increased(p<0.01) compared with control group($2.3{\pm}1.3$). 6. The number of corpora lutea in GBT treated group($6.1{\pm}3.9$) was significantly increased(p<0.01) compared with control group($1.6{\pm}2.3$). 7. The expression of NGF-immunoreactive cells in the ovarian granulosa cells in GBT treated group was lesser observed than control group. Conclusion: From the above results, we concluded that Gwibi-Tang has inhibitory effect on the development of EV-induced polycystic ovary. And it's effect may be related with decreased NGF activities in the ovary.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.1
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• pp.32-39
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• 2018

Objective: In this study, we investigated the adverse effects of dietary zearalenone (ZEA) (0.5 to 1.5 mg/kg diet) on the localization and expression of the growth hormone receptor (GHR) in the uteri of post-weaning gilts and explored alternative mechanism of the reproductive toxicity of ZEA on piglets. Methods: A total of forty healthy piglets (Duroc${\times}$Landrace${\times}$Large White) aged 28 d were selected for study. Piglets were transferred to single cages after 10 days' adaptation on an obstetric table. The animals were allocated to one of four treatments: a normal basal diet supplemented with 0 (Control), 0.5 (ZEA0.5), 1.0 (ZEA1.0), or 1.5 (ZEA1.5) mg/kg purified ZEA, and fed for 35 d after the 10-d adaptation. Analyzed ZEA concentrations in the diets were 0, $0.52{\pm}0.07$, $1.04{\pm}0.03$, and $1.51{\pm}0.13mg/kg$, respectively. At the end of the feeding trial, piglets were euthanized after being fasted for 12 h. Two samples of uterine tissue from each pig were rapidly collected, one of which was stored at $-80^{\circ}C$ for analysis of the relative mRNA and protein expression of GHR, and the second was promptly fixed in Bouin's solution for immunohistochemical analysis. Results: The relative weight of the uteri and thickness of the myometrium and endometrium increased linearly (p<0.001) and quadratically (p<0.001) with an increasing level of ZEA. The results of immunohistochemical analysis indicated that GHR immunoreactive substance was mainly localizated in the cytoplasm of uterine smooth muscle, glandular epithelial, luminal epithelial, stromal, and vascular endothelial cells. In contrast, nuclear staining was rarely observed. The immunoreactive integrated optic density of GHR in the myometrium, luminal epithelium, glandular epithelium, and whole uteri of weaning gilts increased linearly (p<0.001) and quadratically (p<0.05) with an increasing level of ZEA. The mRNA and protein expression of GHR in the uteri of weaning gilts increased linearly (p<0.001) and quadratically (p<0.05) with an increasing level of ZEA. Conclusion: In conclusion, ZEA at a concentration of 0.5 mg/kg was sufficient to significantly thicken the myometrium and endometrium, and at a concentration of 1.0 mg/kg induced a high level of GHR expression to promote growth and development of the uteri. This revealed an alternative molecular mechanism whereby ZEA induces growth and development of the uteri and provides a theoretical basis for the revision of Chinese feed hygiene standards.

• Clinical and Experimental Pediatrics
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• v.45 no.12
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• pp.1551-1558
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• 2002

Purpose : Loss of hippocampal interneurons in dentate gyrus has been reported in patients with severe temporal lobe epilepsy and in animals treated with kainic acid(KA). Interneurons contain $Ca^{2+}$- binding protein parvalbumin(PV). The effects of kainic acid on parvalbumin-immunoreactive (PV-IR) interneurons in dentate gyrus were investigated in organotypic hippocampal slice cultures. Methods : Cultured hippocampal slices from postnatal day nine C57/BL6 mice were exposed to $10{\mu}M$ KA, and were observed at 0, 8, 24, 48, 72 hours after a one hour KA exposure. Neuronal injury was determined by morphologic changes of PV-IR interneuron in dentate gyrus. Results : Transient(1 hour) exposure of hippocampal explant cultures to KA produced marked varicosities in dendrites of PV-IR interneuron in dentate gyrus and the shaft of interbeaded dendrite is often much thinner than those in control. The presence of varicosities in dendrites was reversible with KA washout. The dendrites of KA treated explants were no longer beaded at 8, 24, 48 and 72 hours after KA exposure. The number of cells in PV-IR interneurons in dentate gyrus was decreased at 0, 8 hours after exposure. But there was no significant difference in 24, 48 and 72 hours recovery group compared with control group. Conclusion : The results suggested that loss of PV-IR interneurons in dentate gyrus is transient, and is not accompanied by PV-IR interneuronal cell death.

• Tuberculosis and Respiratory Diseases
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• v.41 no.5
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• pp.462-474
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• 1994

Background : In acute lung injury, activated neutrophils play an important role in tissue damage. For neutrophils to participate in lung inflammation, chemotactic factors released from mononuclear phagocytes are needed to bring these cells to the local site of inflammation, with interleukin-8 (IL-8) being one of the most specific and important chemotactic factors for neutrophils. IL-8 also induces the expression of adhesion molecules and activates neutrophils to release various inflammatory mediators. Lipopolysaccharide(LPS) is one of the most important causes of adult respiratory distress syndrome and can cause release of many inflammatory cytokines including IL-8 leading to acute lung injury. But little is known about the regulatory mechanism of LPS-induced IL-8 gene expression in mononuclear phagocytes. Method : Human alveolar macrophages(HAM) and peripleral blood monocytes(PBMC) were isolated from healthy volunteers. Time and dose relationship of LPS-induced IL-8 mRNA expression was observed by Northern blot analysis. To evaluate the regulatory mechanism of LPS-induced IL-8 gene expression, pretreatment of actinomycin D(AD, $5{\mu}g/ml$) and cycloheximide(CHX, $5{\mu}g/ml$) was done and Northern blot analysis for IL-8 mRNA and ELISA for immunoreactive IL-8 protein in culture supernatant were performed. Results : 1) In HAM, dose and time dependent LPS-induced IL-8 mRNA expression was observed with peak mRNA level at 8 hours post-stimulation. 2) In PBMC, dose and time dependent LPS-induced IL-8 mRNA expression was also observed with peak mRNA level at 4 hours post-stimulation. 3) AD decreased expression of LPS-induced IL-8 gene expression at both mRNAand protein levels in both types of cells. 4) CHX decreased expression of LPS-induced IL-8 gene expression at protein level in both cell types but in HAM, superinduction of IL-8 mRNA was observed while decreased expression of IL-8 mRNA was observed in PBMC. Conclusion : Time and dose dependent LPS-induced IL-8 gene expression was observed in mononuclear phagocytes which is at least partly regulated pretranslationally. LPS-induced IL-8 mRNA expression in HAM needs no de novo protein synthesis and may be under the control of a labile repressor protein while de novo protein synthesis may be needed in PBMC.

• Applied Microscopy
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• v.24 no.2
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• pp.78-92
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• 1994

Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.3
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• pp.185-195
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• 2000

Vascular diseases are significant complications of diabetes mellitus (DM), and the endothelial cells may play a pivotal role in the development of vascular disease in DM. Endothelin-1 (ET-1) released from endothelium is a potent vasoconstrictor peptide and circulating level of ET-1 is increased in a variety of disease states. The purpose of this study was to determine the changes of responsiveness to ET-1 in DM, and we experimented on the changes in the ET-1-induced contraction, levels of nitrite and lipid peroxidation, and ET-1 immunoreactivity in aorta from streptozotocin-induced DM rats. DM was induced by single injection of streptozotocin (55 mg/kg, i.p.). The immunoreactive ET-1 levels in endothelial layer of thoracic aorta were much higher in DM rats than control rats. Nitrite in tissue homogenate was decreased and plasma nitrite was increased in DM rats. Malondialdehyde (MDA) was significantly increased in DM rats and cGMP was not significantly different between control and DM rats. ET-1 produced concentration- dependent contractile responses that are significantly attenuated in DM rats compared to controls. In the presence of selective $ET_A$ receptor antagonist BQ610, the maximum contraction was decreased and the concentration ratios for BQ610 yielded $pA_2$ values of 7.3 (slope, 0.65) in control rats, whereas BQ610 had no antagonistic effect on ET-1-induced contraction in DM rats. However, pretreatment with BQ788, an $ET_B$ receptor antagonist, maximum response was decreased and the dose-response curves for ET-1 were shifted to the right in both groups and $pA_2$ values were 7.9 and 7.7 (slope, 1.05 in control and DM rats), respectively. IRL 1620 and sarafotoxin S6c, $ET_B$ agonists, induced relaxation in control rats but not in DM rats. These results indicate that endothelial cell dysfunction and enhanced immunoreactivity of ET-1 have been found in DM rat and ET-1-induced contraction was attenuated in DM rat. These attenuated responses might be at least in part caused by the alteration of $ET_A$ receptor properties (e.g. desensitization), and partly related with an alteration in intracellular mechanism for contraction to ET-1.