• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.3
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• pp.177-185
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• 2010

Introduction: Epidermal growth factor receptor (EGFR) is expressed in human epithelial tumors including salivary cancers, and known to be correlated with tumor progression and poor clinical courses in some epithelial tumors. In this study, we determined the therapeutic effect of the anti-EGFR monoclonal antibody Erbitux (C225, cetuximab) in combination with the DNA topoisomerase I inhibitor irinotecan (CPT-11) on human salivary adenoid cystic carcinoma (SACC) cells growing in nude mice. Materials and Methods: At first, immunocytochemical analysis for the expression of EGFR and phosphorylated EGFR (pEGFR) on a human salivary ACC cell line (ACC3). To determine the in vivo effects of Erbitux and CPT-11, nude mice with orthotopic parotid tumors were randomized to receive intraperitoneal Erbitux (1 mg) two times per week, intraperitoneal Irinotecan (50 mg/kg) once per week, Erbitux plus CPT-11, or placebo. (control) Tumor volume and weight were measured. And mechanisms of in vivo activity of Erbitux and/or CPT-11 were determined by immunohistochemical/ immunofluorescent analyses. Results: Immunocytochemical staining of ACC3 demonstrated that EGFR was expressed and phosphorylated. CPT-11 inhibited ACC tumor growth in nude mice. Tumors of mice treated with CPT-11 and CPT-11 plus Erbitux exhibited increased tumor cell apoptosis and decreased microvessel density, which correlated with a decrease in the tumor volume in nude mice. But, CPT-11 seems not to be synergistic with Erbitux in our ACC3 model system. Conclusion: These results suggest that anti-EGFR monoclonal antibody and the DNA topoisomerase I inhibitor will be effective in the treatment of recurred or metastatic lesions of salivary ACC.

• Nutrition Research and Practice
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• v.15 no.3
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• pp.319-328
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• 2021

BACKGROUND/OBJECTIVES: Curcuma zedoaria R. (Zingiberaceae) has been used to treat headache, fever, and hypertension-related symptoms in Asian countries, including Korea, China, and Japan. We investigated whether dietary intake of a C. zedoaria extract (CzE) affected atherosclerosis in vivo. MATERIALS/METHODS: Apolipoprotein E-deficient (ApoE^-/-) mice (n = 32) were fed a normal diet (ND), a high-cholesterol diet (HCD), an HCD containing CzE (100 mg/kg/day), or an HCD containing simvastatin (10 mg/kg/day) for 12 weeks. The anti-atherosclerotic effects were evaluated by observing changes in fatty streak lesions, immunohistochemical analysis, ex vivo fluorescence imaging, lipid profiles, and western blot analysis. RESULTS: The CzE-fed group showed a 41.6% reduction of atherosclerosis. Furthermore, CzE significantly reduced the levels of serum triglyceride, high-density lipoprotein, the chemokine (C-X3-C-motif ) ligand 1, the adhesion molecules vascular cell adhesion molecule-1, intracellular adhesion molecule-1, and E-selectin; down-regulation of tumor necrosis factor-α, interleukin-6, high mobility group box-1, and cathepsin levels in the aortic sinuses and aortas of ApoE^-/- mice were also observed. CONCLUSIONS: The results suggest that the inclusion of a water extract of C. zedoaria in a HCD is closely correlated with reducing the risk of vascular inflammatory diseases in an ApoE mouse model.

• Journal of the korean academy of Pediatric Dentistry
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• v.21 no.2
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• pp.439-455
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• 1994

The purpose of this study was to investigate the distribution of nerves in the periodontal ligament of a dog's primary teeth by each developing stage. The distribution of nerves in the periodontal ligament were investigated by means of immunohistochemistry for detection of neurofilament protein (NFP). The results were as follows: The NFP-immunoreactive nerve fibers were found to be densely distributed in the apical third of the periodontal ligament, while they were sparse in the coronal two third, in both primary and permanent teeth. In generally the density of distribution and degree of arborization of nerve fibers in periodontal ligament of primary teeth revealed a poor appearance compared with that of permanent teeth. Periodontal ligament in anterior teeth showed more abundant nerve innervation than posterior teeth, and the periodontal ligament of the bifurcation area in posterior teeth roots were not observed to have nerve fiber. The density of nerve distribution in the periodontal ligament of primary teeth was reduced according to the physiological root resorption and nerve fibers were not observed in the surrounding area on the root of the exfoliation stage in primary teeth. The distribution of nerve fibers in mucogingival tissue, was poor innervated according to the aging of the dogs. A more abundant distribution of nerve fiber was represented in the lingual mucogingival tissue than in the labial side. Most of the nerve endings in the periodontal ligament of primary teeth showed a tree-like appearance. However, the typical Ruffini-like nerve endings were not observed.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.121-130
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• 1994

Expression of gonadotropin releasing hormone(GnRH) has been described in the rat ovary. It remains, however, unkown whether GnRH is synthesized as a prohormone. Therefore, this study was performed to verify the expression of pro-GnRH by in situ hybridization and further to investigate the effect of gonadotropin on GnRH or GnRH mRNA in rat ovary by immunohistochemical and in situ hybridization techniques. Adult female Sprague-Dawely rats were used and the estrous cycle was synchronized by intraperitoneal injection of pregnant mare's serum gonadotropin(PMSG). Ovaries were fixed with 4% paraformaldehyde and embedded with G.C.T. compound and cut by cryostat. For immunohistochemistry, avidin-biotin peroxidase complex(ABS) method was employed and for in situ hybridization, $^{35}S$-end labeled oligonucleotide was used and followed by autoradiography. By in situ hybridization using GnRH oligomer and GAP(GnRH associated protein) oligomer, GnRH mRNA and GAP mRNA were co-localized in the fullicular cells, luteal cells, interstitial cells and theca cells. GnRH or GnRH mRNA signals in the ovary increased by human chorionic gonadotropin(hCG) injection. At the 3 and 6 hrs after hCG injection, the number of GnRH and GnRH mRNA containing cells increased rapidly and the density of GnRH and GnRH mRHA culminated at 9 hrs after heG injection. With the follicular development, the high expression of GnRH and GnRH mRNA was also observed within the follicles. After ovulation, the density of GnRH or GnRH mRNA decreased in the follicles but increased in the corpus lutea.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.6
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• pp.2277-2281
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• 2015

Objectives: To explore the expression of astrocyte elevated gene-1 (AEG-1) in cervical cancer and analyze its correlation with microvascular density (MVD), nuclear factor kappaB (NF-kB p65) and vascular endothelial growth factor (VEGF). Materials and Methods: Immunohistochemical MaxVision method was adopted to detect the expression level of AEG-1, NF-kB p65 and VEGF in 45 samples of invading cervical cancer and 12 samples of cervicitis from The First Affiliated Hospital of Wenzhou Medical University. Tumor microvascular endothelial marker CD34 combined with Weidner was used to determine the MVD in cervical cancer tissue. The positive expression and staining conditions of AEG-1, NF-kB p65 and VEGF in cervical cancer tissues were observed under a light microscope. Correlations between expression of AEG-1 protein and those of NF-Kb p65 and VEGF, as well as MVD, were analyzed using Pearson correlation. Results: The expression levels of AEG-1 were $0.186{\pm}0.043$ in cervical cancer and $0.051{\pm}0.002$ in chronic cervicitis (p<0.01). Moreover, expression of AEG-1 was related to vascular invasion and lymphatic metastasis of cervical cancer (p<0.01), but not with age of the patients, differentiation degree, tumour size, pathological type and parametrial infiltration (p>0.05). Pearson correlation analysis showed that the expression of AEG-1 was linked with NF-kB p65 (r=0.501, p=0.000), VEGF (r=0.718, p=0.000) as well as MVD in cervical cancer tissue (r=0.815, p=0.000). Conclusions: AEG-1 is highly expressed in cervical cancer and promotes angiogenesis, which might be related to the fact that AEG-1 activating the signal pathway of NF-kB could up-regulate the level of VEGF expression.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.5
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• pp.1116-1124
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• 2009

To determine whether topical application of trimix extracts of Mylabris phalerata Pall., Arisaematis Rhizoma and Pinelliae Rhizoma Ternata lead to affects on the hair growth activity in C57BL/6N mice. To examine the hair growth activity of the extracts of Mylabris phalerata Pall., Arisaematis Rhizoma and Pinelliae Rhizoma Ternata gross, microscopic, and immunohistochemical method were performed. In order to examine the mRNA expression of hair growth related substance, RT-PCR method was performed. Experimental group I on day 14, The most extensive hair growth activity was observed in whole skin area of all the mice whose hair had been clipped. Brdu immunoreactive cells of all the experimental groups were more heavily stained in epidermis, bulge, outer root sheath, inner root sheath, subcutaneous tissue, hair bulb and cutaneous trunci muscles than that of control group on day 12 of hair growing cycle in C57BL/6N mice. VEGF immunoreactive density of all the experimental groups was more heavily stained in epidermis, bulge and cutaneous trunci muscles than that of control group on day 12. FGF and c-kit immunoreactive cells of all the experimental groups were heavily stained in epidermis, outer root sheath, inner root sheath and cutaneous trunci muscles on day 12. PKC-$\alpha$ immunoreactive density of all the experimental groups was mildly stained in epidermis and cutaneous trunci muscles than that of control group on day 12. On day 12, the expression of bFGF (138%, 119%, 120%), VEGF (146%, 144%, 133%), IGF-1 (165%, 141%, 119%) and PLI (121%, 116%, 123&) in each experimental groups was more increased than that of control group. On day 16, The expression of IGF-1 (126%, 149%, 151%) in all the experimental group was more increased than that of control group (100%). The expression of bFGF (92%, 94%) and VEGF (101%, 97%), PL1 (102%, 109%) in all the experimental group was more decreased than that of experimental group I, II on day 12. But the expression of bFGF (109%) and VEGF (127%), and PL1 (105%) in each experimental group III was more increased than that of control group (100%). These experiments suggest that trimix extracts of Mylabris phalerata pall., Arisaematis Rhizoma and Pinelliae Rhizoma Ternata may stimulate the topical hair growth activity and its experimental group I can be useful for treatment of alopecia areata.

• Archives of Plastic Surgery
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• v.42 no.6
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• pp.686-694
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• 2015

Background Rosa damascena, a type of herb, has been used for wound healing in Eastern folk medicine. The goal of this study was to evaluate the effectiveness of rose placenta from R. damascena in a full-thickness wound model in mice. Methods Sixty six-week-old C57BL/6N mice were used. Full-thickness wounds were made with an 8-mm diameter punch. Two wounds were made on each side of the back, and wounds were assigned randomly to the control and experimental groups. Rose placenta ($250{\mu}g$) was injected in the experimental group, and normal saline was injected in the control group. Wound sizes were measured with digital photography, and specimens were harvested. Immunohistochemical staining was performed to assess the expression of epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), transforming growth factor-${\beta}1$ (TGF-${\beta}1$), and CD31. Vessel density was measured. Quantitative analysis using an enzyme-linked immunosorbent assay (ELISA) for EGF was performed. All evaluations were performed on postoperative days 0, 2, 4, 7, and 10. Statistical analyses were performed using the paired t-test. Results On days 4, 7, and 10, the wounds treated with rose placenta were significantly smaller. On day 2, VEGF and EGF expression increased in the experimental group. On days 7 and 10, TGF-${\beta}1$ expression decreased in the experimental group. On day 10, vessel density increased in the experimental group. The increase in EGF on day 2 was confirmed with ELISA. Conclusions Rose placenta was found to be associated with improved wound healing in a mouse full-thickness wound model via increased EGF release. Rose placenta may potentially be a novel drug candidate for enhancing wound healing.

• Journal of the korean academy of Pediatric Dentistry
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• v.26 no.4
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• pp.688-702
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• 1999

The purpose of this study was to investigate the revascularization and reinnervation of calcitonin gene-related peptide immunoreactive nerves in immediately replanted rat molars. First maxillary right molars in 56 rats(35days old) were extracted and immediately replanted. The rats were killed 1, 2, 3, 7, 14, 28 and 42 days after replantation and revascularization of pulpal blood vessels were examined microangiogram with korean traditional ink and reinnervation of pulpal nerve were examined immunohistochemical method using calcitonin gene-related peptide(CGRP) antiserum. The results were as follows; 1. Revascularization and reinnervation of CGRP immunoreactive nerve fibers were observed mesial side whole pulp tissue of replanted teeth. Revascularization and reinnervation of CGRP immunoreactive fiber were made at 2days after replantation in entire pulp of replanted teeth and the distribution density of blood vessels were gradually increased according time elapsed, but did not achieve the density of control. 2. Postoperative dentin formation in replanted teeth revealed at 1week after replantaton and gradually increased according to time elapsed. 3. Revascularization and Reinnervation of CGRP immunoreactive nerve fibers were established at the same time and it seems to be closed relatationship between revascularization and reinnervation.

• 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.

• Journal of Gastric Cancer
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• v.20 no.2
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• pp.190-201
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• 2020

Purpose: This study sought to investigate the prognostic significance of tumor-infiltrating lymphocytes (TILs) in relation to tumor location within the stomach. Materials and Methods: The densities and prognostic significance of TIL subsets were evaluated in 542 gastric cancer patients who underwent gastrectomy. Immunohistochemical staining for CD3, CD4, CD8, forkhead/winged helix transcription factor (Foxp3), and granzyme B was performed. Results: Cardia cancer was associated with significantly lower densities of CD8 T-cells and higher densities of Foxp3 and granzyme B T-cells than non-cardia tumors. Multivariate analysis showed that advanced age (hazard ratio [HR], 1.023; 95% confidence interval [CI], 1.006-1.040), advanced T classification (HR, 2.029; 95% CI, 1.106-3.721), lymph node metastasis (HR, 3.319; 95% CI, 1.947-5.658), low CD3 expression (HR, 0.997; 95% CI, 0.994-0.999), and a high Foxp3/CD4 ratio (HR, 1.007; 95% CI, 1.001-1.012) were independent predictors of poor overall survival in cardia cancer patients. In non-cardia cancer patients, total gastrectomy (HR, 2.147; 95% CI, 1.507-3.059), advanced T classification (HR, 2.158; 95% CI, 1.425-3.266), lymph node metastasis (HR, 1.854; 95% CI, 1.250-2.750), and a low Foxp3/CD4 ratio (HR, 0.978; 95% CI, 0.959-0.997) were poor prognostic factors for survival. Conclusions: The densities and prognostic effects of TILs differed in relation to the location of tumors within the stomach. The contrasting prognostic effects of Foxp3/CD4 ratio in cardia and non-cardia gastric cancer patients suggests that clinicians ought to consider tumor location when determining treatment strategies.