• The Journal of Internal Korean Medicine
• /
• v.38 no.6
• /
• pp.891-901
• /
• 2017

Objectives: The aim of the study was to evaluate the anti-asthmatic effect of alismatis rhizoma and alisol acetate B combination therapy in a murine asthma model. Methods: C57BL/6 mice were sensitized to and challenged with a mixture of ragweed, dust mite, and aspergillus to induce an asthma animal model. Alismatis rhizoma extract and alisol acetate B combination therapy was co-administered only in the experimental group. To evaluate the anti-asthmatic effect of the combination therapy, inflammatory cell counts in bronchoalveolar lavage (BAL) fluid were determined, and tissue was examined histologically with hematoxylin and eosin (H & E) and periodic acid-Schiff (PAS) stains, by enzyme-linked immunosorbent assay (ELISA) of IgE, IL-4, and IL-5, and with reverse transcription polymerase chain reaction (RT-PCR) of IL-5, IL-33, MUC5AC. Results: Alismatis rhizoma and alisol acetate B combination therapy reduced the number of inflammatory cells, alleviated histologic features, and down-regulated all the investigated asthma mediators, IgE, IL-4, IL-5, IL-33, and MUC5AC. Conclusions: According to the above results, alismatis rhizoma and alisol acetate B combination therapy may have therapeutic potential for asthma.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.20 no.6
• /
• pp.1593-1597
• /
• 2006

We hope to evaluate the effects of Gami-Choakwiyeum (GCKY) on the PPAR-${\gamma}$’ in the OVA induced asthma mouse model. Female BALB/c mice, 8 weeks of age and free of murine specific pathogens were used. Mice were sensitized by intraperitoneal injection of OVA emulsified in aluminum hydroxide in a total volume of 200 ${\mu}{\ell}$ on one day and 14 days. On 21, 22, and 23 days after the initial intraperitoneal injection of OVA, the mice were challenged using an ultrasonic nebulizer. GCKY was administered 7 times by oral gavage at 24 hour intervals fromdays 19 after intraperitoneal injection of OVA. Bronchoalveolar lavage was perfromed 72 hours after the last challenge, and total cell numbers in the BAL fluid were counted. Also, the level of PPAR-${\gamma}$ of normal and OVA-induced asthma moused with/without administration of GCKY were measured by Western blot analysis. For the histologic examination, the specimens were stained with hematoxylin 2 and eosin-Y.(H & E). Numbers of total cells were increased significantly at 72 h after OVA inhalation compared with numbers of total cells in the normal and the administration of GCKY. Especially, the increased numbers of eosinophils in BAL fluids after OVA inhalation were significantly increased. However, the numbers of eosinophils reduced by the administration of GCKY. Western blot analysis revealed that PPAR-${\gamma}$ levels in nuclear level were increased slightly after OVA inhalation compared with the levels in the normal group. After the administration of GCKY, PPAR-${\gamma}$ levels in cytosolic and nuclear levels at 72 h after OVA inhalation were markedly increased. On pathologic examination, there were many acute inflammatory cells around the alveoli, bronchioles, and airway lumen of mice with OVA-induced asthma compared with inflammatory cells in the normal group. However, acute inflammatory cells around alveoli, bronchioles, and airway lumen markedly decreased after administration of GCKY, GCKY can increase a PPAR-${\gamma}$ level and could be an effective treatment in asthma patients through the PPAR-${\gamma}$ mechanism for bronchial asthma.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.12 no.10
• /
• pp.4378-4384
• /
• 2011

In order to evaluate the anti-oxidative capabilities of Angelica dahurica Radix ethanol extract (ADEE), we analyzed the contents of polyphenol and flavonoid compounds and the electron donating ability from ADEE. For animal experimentation, the test agent was topically applied to the artificial tanning spots which were induced by 1,500 mJ/$cm^2$ of ultraviolet B radiation on the backs of brown guinea pigs weighing approximately 450~500g. The test agent of $30{\mu}{\ell}$ was applied (6areas per group) twice a day, five days a week, for five weeks. On completion of the experiment, the animals were sacrificed under anesthetization, and the artificial tanning spots were obtained by biopsy punch and stained with H&E to observe the histological change in the epidermis and dermis. As a result, the contents of polyphenol and flavonoid compounds in ADEE were 20.7mg/g and 19.5mg/g respectively. As the for electron-donating capability of ADEE, it was observed that ADEE displays a dose-dependent antioxidative capacity of 14.8% and 19.8% at the concentration of 500 and 1000 ${\mu}g/m{\ell}$ respectively. Tissue staining with H&E revealed that the epidermis of the control group was slightly thicker than that of the other groups. However no inflammation or any other undesirable effect on the skin tissue due to ADEE was observed. These results indicate that ADEE is of value as a natural antioxidant.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.1
• /
• pp.63-68
• /
• 2013

Background: Tumor angiogenesis correlates with recurrence and appears to be a prognostic factor for both breast and prostate cancers. In the present study, we aimed to investigate the correlation of microvessel density (MVD), a measure of angiogenesis, with nuclear pleomorphism, mitotic count, and vascular invasion in breast and prostate cancers at preclinical and clinical levels. Methods: Samples from xenograft tumors of luminal B breast cancer and prostate adenocarcinoma, established by BT-474 and PC-3 cell lines, respectively, and commensurate human paraffin-embedded blocks were obtained. To determine MVD, specimens were immunostained for CD-34. Nuclear pleomorphism, mitotic count, and vascular invasion were determined using hematoxylin and eosin (H&E)-stained slides. Results: MVD showed significant correlations with nuclear pleomorphism (r=0.68, P=0.03) and vascular invasion (r=0.77, P=0.009) in breast cancer. In prostate cancer, MVD was significantly correlated with nuclear pleomorphism (r=0.75, P=0.013) and mitotic count (r=0.75, P=0.012). In the breast cancer xenograft model, a significant correlation was observed between MVD and vascular invasion (r=0.87, P=0.011). In the prostate cancer xenograft model, MVD was significantly correlated with all three parameters (nuclear pleomorphism, r=0.95, P=0.001; mitotic count, r=0.91, P=0.001; and vascular invasion, r=0.79, P=0.017; respectively). Conclusions: Our results demonstrate that MVD is correlated with nuclear pleomorphism, mitotic count, and vascular invasion at both preclinical and clinical levels. This study therefore supports the predictive value of MVD in breast and prostate cancers.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.16
• /
• pp.7385-7390
• /
• 2015

Background: Transthoracic fine needle aspiration (FNA) cytology and core needle biopsy (CNB) are two commonly used approaches for the diagnosis of suspected neoplastic intrathoracic lesions. This study compared the diagnostic accuracy of FNA cytology and concurrent CNB in the evaluation of intrathoracic lesions. Materials and Methods: We studied FNA cytology and concurrent CNB specimens of 127 patients retrospectively, using hematoxylin and eosin (H&E), immunohistochemistry, and, on certain occasions cytochemistry. Information regarding additional tissue tests was derived from the electronic archives of the Department of Pathology and Laboratory Medicine as well as patient records. Diagnostic accuracy was calculated for each test. Results: Of 127 cases, 22 were inconclusive and excluded from the study. The remaining 105 were categorized into 73 (69.5%) malignant lesions and 32 (30.5%) benign lesions. FNA and CNB findings were in complete agreement in 63 cases (60%). The accuracy and confidence intervals (CIs) of FNA and CNB for malignant tumors were 86.3% (CI: 79.3-90.7) and 93.2% (CI: 87.3-96.0) respectively. For epithelial malignant neoplasms, a definitive diagnosis was made in 44.8% of cases by FNA and 80.6% by CNB. The diagnostic accuracy of CNB for nonepithelial malignant neoplasms was 83.3% compared with 50% for FNA. Of the 32 benign cases, we made specific diagnoses in 16 with diagnostic accuracy of 81.3% and 6.3% for CNB and FNA, respectively. Conclusions: Our findings suggest that FNA is comparable to CNB in the diagnosis of malignant epithelial lesions whereas diagnostic accuracy of CNB for nonepithlial malignant neoplasms is superior to that for FNA. Further, for histological typing of tumors and examining tumor origin, immunohistochemical work up plays an important role.

• Development and Reproduction
• /
• v.1 no.2
• /
• pp.189-202
• /
• 1997

The hypothalamic peptide GnRH plays a central role in the regulation of the mammalian reproductive axis. Recent studies suggested that GnRH stimulates or inhibits the ovarian steroidogenesis and gametogenesis directly. Our previous report indicated that GnRH gene is expressed in adult rat ovary as well as in hypothalamus and that the expressed GnRH may induce the follicular atresia and apoptosis of ovarian granulosa cells in rat. Therfore, we studied whether GnRH gene is expressed in the mouse fetal ovary, when the germ cells are degenerating by apoptosis during gonad diffeerentiation. Mouse fetal gonads were obtained on the 12, 15,18 and 20th day of gestation from the mother mice superovulated (10 IU PMSG and 10 IU hCG) and mated. The morphological changes of fetal ovaries were examined histochemically by hematoxylin-eosin staining. The fetal sex was confirmed by PCR methods for sexing. RT-PCR methods were used to examine the expression of GnRH gene and the sex steroid hormones were determined by conventional radioimmunoassays. The levels of estradiol (E) and progesterone (P) were increaseduntil 18th day of gestation and then E was decreased just before parturition. The morphological changes of fetal gonadal tissue sections showed the ovarian development and coincided with the result of PCR analysis for sexing using ovary- or testis- specific oligonucleotide primers. Immunoreactive GnRH in placenta was decreased gradually until the end of gestation but fetal brain and ovarian GnRH were increased. The level of GnRH gene expression was increased during fetal ovarian development from 12 till 18th day and decreased suddenly on 20th day just before birth. From these results, it is suggested that ovarian GnRh may play a regulatory role on the germ cell differentiation of fetal ovary.

• Journal of Korean Medicine Rehabilitation
• /
• v.23 no.1
• /
• pp.25-49
• /
• 2013

Objectives : This study was carried out to know the anti-osteroarthritic effects of Bangkibokryeong-tang(Fanjifuling-tang(BBT)) on the papain-induced osteoarthritis C57BL/10 mouse. Methods : Osteoarthritis was induced by injection of papain(6 ${\mu}l$) into knee joint of mouse. Osteoarthritic mice were divided into 4 groups(normal, control, joins(R), BBT). The injection did not fit the normal group. A week later, after the injection of papain, control group was taken normal saline 200 ${\mu}l$, positive control group was taken joins(R)(100 mg/kg), treated group was taken extract of Bangkibokryeong-tang(Fanjifuling-tang(BBT))(400 mg/kg). After then, we examined hepatotoxicity, nephrotoxicity, inflammation cytokines, expression of inflammation factor mRNA, hemotology, histology through the micro CT-arthrography, and etc. Results : 1. Hepatotoxicity and nephrotoxicity have not expressed. 2. The levels of IL-$1{\beta}$, TNF-${\alpha}$, IL-6, MCP-1, Thromboxane B2, Leukotriene B4, Prostaglandin E2 in serum were significantly decreased. 3. In hematology, the levels of neutrophils and monocytes were significantly decreased. 4. The expression of inflammation factor mRNA like TNF-${\alpha}$ and IL-6, COX-2, iNOS-II were significantly inhibited. 5. In micro CT-arthrography, cartilage volume was less decreased. 6. The degree of osteoarthritis induced damage of joint of BBT group is low in histopathologic observation(hematoxylin&eosin(H&E), Safranin-O). Conclusions : According to this study, BBT has effect of anti-osteoarthritis. Further clinical research for the cartilage protective effect is necessary.

• Journal of Life Science
• /
• v.10 no.2
• /
• pp.1-8
• /
• 2000

This experiment was performed to study the structure of esophageal mucousa and the histochemical properties of glycosaminoglycans of esophageal mucous cells in four teleostean species, i.e., Agramus agramus, Inimicus japonicus, Epinephelus chlorostigma and Helicolenus dactylopterus. To observe the structure of eosphageal mucosa, hematoxylin-eosin(H-E) staining was used. The glycosaminoglycans was stained with PAS(periodic acid schiff), alcain blue(AB) pH 2.5, AB pH 1.0, aldehyde fuchsin(AF) pH 1.7, AF pH 1.0, AB pH 2.5-PAS, AB pH 1.0-PAS and AF pH 1.7-alcian blue pH 2.5. As for the amount and histochemical composition of glycosaminglycans in Agramus agramus, most of mucous secreting columnar cell and mucous cells contain large and moderate amount of neutral glycosaminoglycans. A few of mucous cells having small amount of neutral glycosamino-glycans and minimal amount of sulfated glycosamino-glycans. In Inimicus japonicus, the esophageal mucous cells were composed of most of medium sized and large mucous cells with moderate amount of neutral glycosaminoglycanonly only, a few of medium sized and large mucous cells and most of small mucous cells with considerable amount of neutral glycosaminoglycans and minimal to small amount of nonsulfated glycosaminoglycans, and a feq of small mucous cells with small amount of neutral glycosaminoglycans and minimal amount of sulfated glycosaminoglycans. In Epinephelus chlorostigma, most of medium sized and large mucous cells were mixed small amount of neutral glycosaminoglycans with sulfated glycosaminoglycans, a few of which were contained with moderate or considerable amount of neutral glycosaminoglycans with sulfated glycosaminoglycans, while most of small mucous cells containing considerable amount of neutral glycosaminoglycans and small to moderate or considerable to minimal amount of nonsulfated glycosaminoglycans(sialomucin) a few of which containing a mixture of considerable amount of neutral glycosaminoglycans and considerable amount of nonsulfated glycosaminoglycans(sialomucin)only. In Helicolenus dactylopterus, most of medium sized and large mucous cells, mixing with moderate to considerable amount of neutral glycosaminoglycans and small to moderate amount of sulfated glycosaminoglycans, while most of small mucous cells with considerable amount of nonsulfated glycosaminoglycans (sialomucin), a few of which having a mixture of considerable amount of neutral glycosaminoglycans and considerable or small amount of nonsulfated glycosaminoglycans(sialomucin).

• Journal of Korean Neurosurgical Society
• /
• v.29 no.7
• /
• pp.853-860
• /
• 2000

Objective : The purpose of our experimental study was to analysis the advantages and disadvantages in the reversible and irreversible cerebral ischemic models with rats by staining with Neutral Red(NR) solusion, 2% 2,3,5-triphenyltetrazolium chloride(TTC) and Hematoxylin & Eosin(H & E). Methods : We have measured the range of cerebral infarction in the rat to get a suitable ischemic model along the object of study with and without craniectomy. With craniectomy, 9 rats were sacrificed for irreversible cerebral ischemic model by means of ligation at proximal(group I) and distal(group II), and coagulation at proximal(group III) middle cerebral artery. Also, 6 rats were sacrificed for irreversible(group IV) and reversible(group V) cerebral ischemic model using nylon thread without craniectomy. The sizes of infarction were measured by staining the coronal sections of the brain with NR solusion, TTC and H & E. Results : There are no difference of physiological parameters comparing the each group. Cerebral infarction was not observed in group II, but it's volume was largest in group IV. Disadvantages of craniectomy group(I, II, III) are the long duration of operation and cortical damage by procedure. It's advantage is confirmation of the middle cerebral artery occlusion and cessation of blood flow through the operative microscope. In case of ischemic models using nylon thread (group IV, V), it is hard to identify the interruption or recirculation of blood flow through the middle cerebral artery, but the advantage is the simplicity of operative technique which reduces the operation time and minimizes the cerebral damage due to craniectomy. Therefore, it seems important to set up the reversible and irreversible ischemic models by carefully considering advantages and disadvantages listed above. Conclusion : TTC staining seems to be effective since it reflects the histological damage sufficiently and quickly. It is hoped that researches focused on ischemic penumbra, which became popular recently, will be further carried on with use of NR staining, optical microscope and electron microscope.

• Biomedical Science Letters
• /
• v.8 no.3
• /
• pp.179-184
• /
• 2002

We investigated the morphological changes and TUNEL reaction of apoptotic cells in the liver of D-galactosamine (20 mg/mouse) and lipopolysaccharide (5 $\mu\textrm{g}$/mouse)-treated 30 mice (BALB/c), and in additioa also of apoptotic cells in kidney and spleen. The livers and other some organs of mice at 6, 12, 24, 48 and 72 hrs after treatment were collected and were fixed with 10% neutral formalin and paraffin sections were stained with hematoxylin-eosin or terminal deoxynucleotidly transferase-mediated dUTP nick end labeling (TUNEL) method. Morphological changes in apoptotic hepatocytes were chondensation of nuclei and density of cytoplasms, then the margination and pyknosis of chromatin, the formation of half-moon- or horse-shoe- or ship-like shapes of condensed chromatin mass, lastly formation of apoptotic bodies, disappearance of nuclear envelopes, decrease of stainability, then lysis and disappearance of apoptotic bodies. TUNEL positive reactions of hepatocytes were appeared first moderate in uncondensed hepatocytes, severe in condensed hepatocytes, moderate in chromatin-marginated hepatocytes. These reactions also were appeared moderate in hepatocytes with half-moon- or horse-shoe- or ship-like pyknotic chromatin mass or apoptotic bodies, and mild or negative in hepatocytes with lysed apoptotic bodies or with disappeared nuclear envelopes. Consequently these results suggested that TUNEL positive reactions of hepatocytes appeared at more early stages than appearance of chromatin condensation and disappeared at more early stage than disappearance of histological findings of apoptosis. We also confirmed that the differentiation of apoptotic cells from normal healthy cells of Kupffer cells and vascular endothelial cells in liver, reticular cells and lymphocytes in spleen and epithelial cells of tubules and ducts in kidney was impossible in H-E preparations but was possible in TUNEL preparations.