• Title/Summary/Keyword: immunohistochemical methods

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Immunohistochemical studies on the relationship between pineal body and superior cervical ganglia of the Korean native goat (한국재래산양 송과체와 앞쪽목신경절의 관계규명을 위한 면역조직화학적 연구)

  • Lee, Heungshik S.;Lee, In-Se;Song, Seung-hoon;Yoon, Sung-tae;Hwang, In-koo;Lee, Choong-hyun
    • Korean Journal of Veterinary Research
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    • v.40 no.2
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    • pp.197-211
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    • 2000
  • The pineal body have been known to be affected by superior cervical ganglia, and most of its nerve fibers containing peptidergic neurotransmitters have been considered to be originated from this ganglia. To confirm this relationships, some peptidergic neurotransmitters were identified in both of pineal body and superior cervical ganglia of the Korean native goat, which were divided into two group; breeding season and non-breeding season. The localizations of two catecholamine-synthesizing enzymes; tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH), were investigated by immunohistochemistry in the superior cervical ganglia and the pineal body of adult Korean native goats. Substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and galanin (GAL) were also identified in these organs by immunohistochemical and double immunofluorescent methods. In superior cervical ganglia, immunoreactivities for TH and DBH were confirmed in the same ganglion cells. The immunoreactivites for SP, VIP(only in male), NPY and GAL were identified in both of ganglion cell bodies and nerve fibers in the ganglia. CGRP immunoreactivity, however, was observed only in nerve fibers. Most NPY- and VIP-immunoreactive(IR) ganglion cells also contained TH. SP and TH were colocalized in the cell bodies, but not in the nerve fibers. TH immunoreactivity was shown in almost all of ganglion cells in the superior cervical ganglia. The immunoreactivity for NPY had some seasonal variation and was stronger in breeding season than in non-breeding season. In pineal body, lots of TH-IR fibers were observed throughout the parenchyma including the pineal stalk and most of them also contained DBH. SP- and NPY-IR fibers were also immunostained with TH or DBH. But a few SP- and NPY-IR fibers were not colocalized with TH or DBH. Exceptionally, a bipolar neuron-like cell was observed to be immunostained with NPY in the pineal body. A few CGRP and GAL-IR fibers were observed, while VIP-IR fibers were not present. It is concluded that most TH- and DBH-IR fibers as well as the peptidergic immunoreactive fibers of the pineal body might be originated from the superior cervical ganglia. Some peptidergic immunoreactive fibers, however, might be come from other regions of brain. We also suggest that NPY in pineal body plays a important role for pineal function. The seasonal variation of NPY immunoreactivity indicates that the synthesis and use of NPY may be different between in breeding and non-breeding seasons.

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The Effects of Ohyaksungi-san(Wuyaoshunqi-san) on Cognition and Motor Function Recovery after Ischemic Brain Injury in Rat (오약순기산(烏藥順氣散)이 허혈성 뇌손상 흰쥐의 인지 및 운동기능회복에 미치는 효과)

  • Ryu, Ki-Jun;Choi, Jin-Bong;Shin, Mi-Suk;Kim, Sun-Jong
    • Journal of Korean Medicine Rehabilitation
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    • v.18 no.3
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    • pp.1-17
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    • 2008
  • Objectives : Ohyaksungi-san(Wuyaoshunqi-san) has been used for many years as a treatment for cerebrovascular diseases in Oriental medicine. This study was designed to evaluate the effects of Ohyaksungi-san(Wuyaoshunqi-san) on cognition and motor function recovery after ischemic brain injury, and also the expression of BDNF in hippocampus. Methods : This study was designed with 4 subgroups to evaluate the effects of Ohyaksungi-san(Wuyaoshunqi-san). As control groups, group I has no treatment during 1 week after ischemic brain injury and group II has no treatment during 2 weeks after ischemic brain injury. As experimental groups, group III has been treated with Ohyaksungi-san(Wuyaoshunqi-san) during 1 week after ischemic brain injury and group IV have treated with Ohyaksungi-san(Wuyaoshunqi-san) during 2 week after ischemic brain injury. Each group has been examined by tests as follows, neurological motor behavioral tests, cognitive motor behavior test and histological test. Neurological motor behavior tests consisted of limb placement test, beam-walking test and horizontal wire test. Cognitive motor behavior test was performed by using Morris water maze. In the histological test, TTC(2,3,5-triphenylteterazolium chloride) staining, hematoxylin & eosin staining, and immunohistochemical staining were used. Results : 1. The tests for motor function recovery change had significantly good result in the experimental groups as compared with control groups(p<.05). 2. The Morris water maze test on cognition also had significantly good result in the experimental groups as compared with control groups(p<.05). 3. In the immunohistochemical staining for the expression of BDNF in hippocampus, more immune reaction was investigated in the experimental groups as compared with control groups. Especially group IV has the greatest immune reaction. Conclusions : Ohyaksungi-san(Wuyaoshunqi-san) has good effects on cognition and motor function recovery after ischemic brain injury, and also the expression of BDNF in hippocampus.

Expression of E-cadherin and CD44H in Borrmann Type IV Gastric Cancer (Borrmann 4형 위암에서 E-cadherin 및 CD44H의 발현)

  • Choi Won Yong;Yook Jeong Hwan;Shin Dong Gue;Kim Yong Jin;Kim Jung Sun;Oh Sung Tae;Kim Byung Sik;Park Keon Chun
    • Journal of Gastric Cancer
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    • v.4 no.2
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    • pp.82-88
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    • 2004
  • Purpose: E-cadherin and CD44H have been shown to play a role in the progression and the metastasis of tumors. This study evaluated the clinical correlations between expression of E-cadherin and CD44H and various clinicopathologic factors and the value of expressions of E-cadherin and CD44H as prognostic factors in Borrmann type IV gastric cancer. Materials and Methods: In 122 patients with Borrmann type IV gastric cancer, we performed the immunohistochemical stainings for E-cadherin and CD44H. We analyzed the correlation between the expressions of E-cadherin and CD44H and lymphatic invasion, venous invasion, perineural invasion, histologic type, lymph node metastasis, depth of invasion, stage, and peritoneal dissemination, and survival. Results: There were no correlations between reduced expression of E-cadherin and CD44H and lymphatic invasion, venous invasion, perineural invasion, histologic type, lymph node metastasis, depth of invasion, and stage. However, there was a significant correlation between lymph node metastasis and the lymphatic invasion (P=0.022). There was also a significant correlation between the peritoneal dissemination and CD44H expression (P=0.005). The 5-year survival rate was correlated with CD44H expression expression (P=0.026), peritoneal dissemination (P<0.01), depth of invasion (P<0.01), lymph node metastasis (P<0.01), stage of tumor (P<0.01), and lymphatic invasion (P<0.01). There was no correlation between expression of E-cadherin and survival rate. Conclusion: The expression of CD44H and peritoneal dissemination was correlated. The expression of CD44H was an independent prognostic factor in Borrmann type IV gastric cancer. Further prospective studies with a large number of cases are required.

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Comparison of Immunohistochemical Expression of CBP(cAMP-responsive Element Binding Protein) Transcriptional Co-activator between Premalignant Lesions and Squamous Cell Carcinomas in the Lungs (전암성 폐병변 및 편평상피세포폐암 조직에서 CBP(cAMP-responsive Ele-ment Binding Protein) 전사 공동 활성인자의 면역조직화학적 발현양상의 비교)

  • Shin, Jong Wook;Kim, Jin Soo;Kim, Mi Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.2
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    • pp.165-172
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    • 2007
  • Background: The pathogenesis of lung cancer includes the accumulation of multiple genetic abnormalities. The CREB-binding protein(CBP) is one of several transcriptional co-activators among various sequence-specific DNA-binding transcription factors. CBP is involved in a wide range of cellular activities, such as DNA repair, cell growth, differentiation, and apoptosis that are suspected of contributing to tumorigenesis. The goal of this study was to evaluate CBP expression in a series of human lung tissues containing normal epithelium, premalignant lesions(hyperplasia and dysplasia) and squamous cell carcinomas. Materials and Methods: Immunohistochemical staining was performed on formalin-fixed paraffin-embedded sections by use of a monoclonal anti-CBP antibody. CBP expression was compared in samples from 120 patients with premalignant and malignant histological types including 20 metaplastic specimens, 40 dysplastic specimens, and 60 squamous cell carcinomas in the lung. Results: CBP expression was seen in 35% (7/20) of the metaplastic specimens. 65% (26/40) of the dysplastic specimens, and 70% (42/60) of the squamous cell carcinomas (p<0.05). According to celluar atypism, CBP expression was 50% (10/20) of the low-grade dysplastic specimens and 80% (16/20) of the high-grade dysplastic specimens(p <0.01). By cellular differentiation, CBP expression was seen in 95% (19/20) of the well differentiated squamous cell carcinomas, 85% (17/20) of the moderately differentiated carcinomas and 30% (6/20) of the poorly differentiated lesions (p <0.05). Conclusion: These results suggest that CBP may have an important role in malignant transformation of precancerous lung lesions and may be a marker for malignancy.

Neuroprotective Effects of Sopung-tang(Shufeng-tang) on Cognition and Motor Function Recovery after Ischemic Brain Injury in Rats (소풍탕이 허혈성 뇌손상 흰쥐의 인지 및 운동기능회복에 미치는 효과)

  • Chu, Min-Gyu;Choi, Jin-Bong;Shin, Mi-Suk;Kim, Sun-Jong
    • Journal of Korean Medicine Rehabilitation
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    • v.18 no.2
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    • pp.45-60
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    • 2008
  • Objectives : Sopung-tang(Shufeng-tang) is a famous herbal prescription that treated ischemic brain injury. This study was designed to evaluate the effects of Sopung-tang(Shufeng-tang) on congnition and motor function recovery after ischemic brain injury in rats. Methods : Male rats were divided into 4 groups. Those rats caused ischemic brain injury by occlusion of MCA as Longa method. Control group I was per os normal saline for 7 days after ischemic brain injury. Control group II was per os normal saline for 14 days after ischemic brain injury. Experimental group I(Ex I) was taken with Sopung-tang(Shufeng-tang) for 7 days after ischemic brain injury. Experimental group II(Ex II) was taken with Sopung-tang(Shufeng-tang) for 14 days after ischemic brain injury. The author carried out neurological, cognitive motor behavior tests and histological assessment. Neurological motor behavior tests consist of limb placement test, beam-walking test and horizontal wire test. Morris water maze test was used for cognitive motor behavior test. In the histological assessment test, TTC(2,3,5-triphenylteterazolium chloride) staining, Hematoxylin & Eosin staining and immunohistochemical staining were experimented. Results : 1. In neurological motor behavior tests, motor function recovery was significantly increased in the experimental groups as compared with control groups(p<0.05). Especially Ex II was significantly increased as compared with Ex I(p<0.05). 2. In Morris water maze test, congnitive motor function recovery was significantly increased in the experimental groups as compared with control group(p<0.05). Especially Ex II was significantly increased as compared with Ex I(p<0.05). 3. In the immunohistochemical staining for the expression of BDNF in hippocampus, more immune reaction was investigated in the experimental groups as compared with control groups. Especially most immune reaction was experimented in the EX II. Conclusions : According to the above results, Sopung-tang(Shufeng-tang) can treat on the congnition and motor function recovery after ischemic brain injury in rats. And it is effective method in expression of BDNF in hippocampus.

Effects of Dokwal-tang(Duhuo-tang) and Jungsongouhyul Pharmacopuncture on Pain Control and Nerve Regeneration after Crush Injury in Rat Sciatic Nerve (흰쥐 좌골신경 압좌 손상 후 독활탕(獨活湯)및 중성어혈약침(中性瘀血藥鍼)이 통증 억제와 신경 재생에 미치는 효과)

  • Song, Jeong-Seob;Na, Cheol;Shin, Byung-Cheul;Lee, Su-Kyung;Kwon, Young-Dal;Song, Yung-Sun
    • Journal of Korean Medicine Rehabilitation
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    • v.18 no.2
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    • pp.61-79
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    • 2008
  • Objectives : The purpose of this study was to identify the effect of Dokwal-tang(Duhuo-tang) and Jungsongouhyul Pharmacopuncture on pain control and nerve regeneration after crush injury in rat sciatic nerve. Methods : Animal model was produced through right sciatic nerve crush injury and they were randomly divided into four groups; Group I: no treatment control group; Group II: experimental group treated with Dokwal-tang(Duhuo-tang); Group III: experimental group treated with Jungsongouhyul Pharmacopuncture; Group IV: experimental group treated with Dokwal-tang(Duhuo-tang) and Jungsongouhyul Pharmacopuncture. For the pain assessment, this study was observed the paw withdrawal latency(PWL) and immunoreactivity on the substance-P. For the nerve regeneration assessment, the sciatic functional index(SFI) and immunoreactivity on the GAP-43 were measured. Results : 1. In the pain assessment, the PWL of experimental groups were significantly higher than control group and group IV was significantly higher than other groups. 2. In immunohistochemical response of substance-P, as time passes, all groups had decreased immunoreactivity gradually. Specially, group IV was observed the lowest immunoreactivity. 3. In the assessment of SFI, the SFI of experimental groups were significantly higher than control group. 4. In immunohistochemical response of GAP-43, all groups had higher GAP-43 immunoreactivity at the 14 days from post-injury and at the all days, control group was observed most lower immunoreactivity and group IV was observed most higher immunoreactivity. Conclusions : These results of this study suggest that Dokwal-tang(Duhuo-tang) and Jungsongouhyul Pharmacopuncture was related with pain inhibition and motor nerve recovery, and decreased substance-P expression and increased GAP-43 expression after sciatic nerve crush injury.

Ultrastructural and Immunohistochemical Study of Hepatic Fibrosis after the Ligation of the Common Bile Duct in Rats (백서의 총담관 결찰에 의한 간 섬유화의 초미세구조적 및 면역조직화학적 연구)

  • Moon, Kyung-Rye;Rho, Young-Ill;Seo, Woo-Chul;Park, Yeong-Bong;Kim, Man-Woo;Seo, Jae-Hong;Park, Sang-Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.2 no.2
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    • pp.185-193
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    • 1999
  • Purpose: Proliferation of bile duct-like structures and fibrosis is a hepatic cellular reaction observed in most forms of human liver disease and in a variety of experimental conditions associated with liver injury. The aim of this study was to investigate the activation of Ito cells and bile duct proliferation in the rat after common bile duct ligation (CBDL). Methods: Hepatic morphological abnormalities were examined in rats whose bile ducts had been irreversibly ligated for 15, 21, 24 and 28 days. The liver was examined by immunohistochemical staining for ${\alpha}$-smooth muscle actin, the known marker of activated Ito cells, and light and electron microscopes. Results: After CBDL, the bile canalicular proliferation and interstitial fibrosis were gradually increased in the periportal areas extended to hepatic sinusoids. Ito cells positive for ${\alpha}$-smooth muscle actin were frequently observed in the periductular space and in perisinusoidal space of Disse. Ito cells and myofibroblasts were gradually increased in the interstitial fibrosis until the 28th day after CBDL. Ito cells and myofibroblasts had microfilaments with dense body at the periphery of the cell. Conclusions: Our results suggest that Ito cells may be fibroblastic or myogenic. It has also been postulated that during the development of hepatic fibrosis, Ito cells become myofibroblasts or fibroblast like cells.

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CDC6 mRNA Expression Is Associated with the Aggressiveness of Prostate Cancer

  • Kim, Ye-Hwan;Byun, Young Joon;Kim, Won Tae;Jeong, Pildu;Yan, Chunri;Kang, Ho Won;Kim, Yong-June;Lee, Sang-Cheol;Moon, Sung-Kwon;Choi, Yung-Hyun;Yun, Seok Joong;Kim, Wun-Jae
    • Journal of Korean Medical Science
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    • v.33 no.47
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    • pp.303.1-303.10
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    • 2018
  • Background: Cell division cycle 6 (CDC6) is an essential regulator of DNA replication and plays important roles in the activation and maintenance of the checkpoint mechanisms in the cell cycle. CDC6 has been associated with oncogenic activities in human cancers; however, the clinical significance of CDC6 in prostate cancer (PCa) remains unclear. Therefore, we investigated whether the CDC6 mRNA expression level is a diagnostic and prognostic marker in PCa. Methods: The study subjects included 121 PCa patients and 66 age-matched benign prostatic hyperplasia (BPH) patients. CDC6 expression was evaluated using real-time polymerase chain reaction and immunohistochemical (IH) staining, and then compared according to the clinicopathological characteristics of PCa. Results: CDC6 mRNA expression was significantly higher in PCa tissues than in BPH control tissues (P = 0.005). In addition, CDC6 expression was significantly higher in patients with elevated prostate-specific antigen (PSA) levels (> 20 ng/mL), a high Gleason score, and advanced stage than in those with low PSA levels, a low Gleason score, and earlier stage, respectively. Multivariate logistic regression analysis showed that high expression of CDC6 was significantly associated with advanced stage (${\geq}T3b$) (odds ratio [OR], 3.005; confidence interval [CI], 1.212-7.450; P = 0.018) and metastasis (OR, 4.192; CI, 1.079-16.286; P = 0.038). Intense IH staining for CDC6 was significantly associated with a high Gleason score and advanced tumor stage including lymph node metastasis stage (linear-by-linear association, P = 0.044 and P = 0.003, respectively). Conclusion: CDC6 expression is associated with aggressive clinicopathological characteristics in PCa. CDC6 may be a potential diagnostic and prognostic marker in PCa patients.

Adjuvant therapy with 1% alendronate gel for experimental periodontitis treatment in rats

  • de Campos Kajimoto, Natalia;de Paiva Buischi, Yvonne;Loomer, Peter Michael;Bromage, Timothy G.;Ervolino, Edilson;Fucini, Stephen Enrico;Pola, Natalia Marcumini;Pirovani, Beatriz Ommati;Morabito, Maria Juliana Sismeiro;de Almeida, Juliano Milanezi;Furlaneto, Flavia Aparecida Chaves;Nagata, Maria Jose Hitomi
    • Journal of Periodontal and Implant Science
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    • v.51 no.6
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    • pp.374-385
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    • 2021
  • Purpose: The aim of this study was to evaluate the effects of locally delivered 1% alendronate (ALN) gel used as an adjunct to non-invasive periodontal therapy. Methods: Ligature-induced periodontitis was performed in 96 rats. The ligature was tied in the cervical area of the mandibular left first molar. The animals were randomly divided into 4 groups: 1) NT, no treatment; 2) SRP, scaling and root planning; 3) SRP/PLA, SRP followed by filling the periodontal pocket with placebo gel (PLA); and 4) SRP/ALN, SRP followed by filling the periodontal pockets with 1% ALN gel. Histomorphometric (percentage of bone in the furcation region [PBF]) and immunohistochemical (receptor activator of nuclear factor-κB ligand, osteoprotegerin, and tartrate-resistant acid phosphatase) analyses were performed. Data were statistically analyzed, with the threshold of statistical significance set at P≤0.05. Results: The SRP, SRP/PLA, and SRP/ALN groups presented a higher PBF than the NT group (P≤0.01) at 7, 15, and 30 days. The SRP/ALN group presented a higher PBF than the SRP/PLA group in all experimental periods, as well as a higher PBF than the SRP group at 15 and 30 days. No differences were observed in the immunohistochemical analyses (P>0.05 for all). Conclusions: Locally delivered 1% ALN gel used as an adjunct to SRP enhanced bone regeneration in the furcation region in a rat model of experimental periodontitis.

Zearalenone regulates key factors of the Kelch-like erythroid cell-derived protein with CNC homology-associated protein 1-nuclear factor erythroid 2-related factor 2 signaling pathway in duodenum of post-weaning gilts

  • Cheng, Qun;Jiang, Shu zhen;Huang, Li bo;Yang, Wei ren;Yang, Zai bin
    • Animal Bioscience
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    • v.34 no.8
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    • pp.1403-1414
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    • 2021
  • Objective: This study explored the mechanism of the Kelch-like erythroid cell-derived protein with CNC homology-associated protein 1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway under conditions of zearalenone (ZEA)-induced oxidative stress in the duodenum of post-weaning gilts. Methods: Forty post-weaning gilts were randomly allocated to four groups and fed diets supplemented with 0, 0.5, 1.0, or 1.5 mg/kg ZEA. Results: The results showed significant reductions in the activity of the antioxidant enzymes total superoxide dismutase and glutathione peroxidase and increases the malondialdehyde content with increasing concentrations of dietary ZEA. Immunohistochemical analysis supported these findings by showing a significantly increased expression of Nrf2 and glutathione peroxidase 1 (GPX1) with increasing concentrations of ZEA. The relative mRNA and protein expression of Nrf2, GPX1 increased linearly (p<0.05) and quadratically (p<0.05), which was consistent with the immunohistochemical results. The relative mRNA expression of Keap1 decreased linearly (p<0.05) and quadratically (p<0.05) in the duodenum as the ZEA concentration increased in the diet. The relative mRNA expression of modifier subunit of glutamate-cysteine ligase (GCLM) increased quadratically (p<0.05) in all ZEA treatment groups and the relative mRNA expression of quinone oxidoreductase 1 (NQO1) catalytic subunit of glutamate-cysteine ligase decreased linearly (p<0.05) and quadratically (p<0.05) in the ZEA1.0 group and ZEA1.5 group. The relative protein expression of Keap1 and GCLM decreased quadratically (p<0.05) in the duodenum as the ZEA concentration increased in the diet, respectively. The relative protein expression of NQO1 increased linearly (p<0.05) and quadratically (p<0.05) in all ZEA treatment groups in the duodenum. Conclusion: These findings suggest that ZEA regulates the expression of key factors of the Keap1-Nrf2 signaling pathway in the duodenum, which enables resistance to ZEA-induced oxidative stress. Further studies are needed to examine the effects of ZEA induced oxidative stress on other tissues and organs in post-weaning gilts.