• Title/Summary/Keyword: Immuno histochemistry

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The Effect of Acupuncture in Promoting Neurogenesis and Angiogenesis after Middle Cerebral Artery Occlusion in Rats

  • Lee, Hong Min;Nam, Sang Soo;Kim, Yong Suk
    • Journal of Acupuncture Research
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    • v.30 no.3
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    • pp.1-13
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    • 2013
  • Objectives : This study was performed to choose more effective neuro-protective acupuncture point and to verify the effect of acupuncture in promoting neurogenesis and angiogenesis as a result of its neuro-vasculo-regenerative effect in middle cerebral artery occlusion model in rats. Methods : By TTc staining we chose the most effective acupuncture point with neuro-protection. We randomly divided into four groups: Such as (1) sham group(with sham-operation), (2) sham+acupuncture group(with sham-operation), (3) middle cerebral artery occlusion group, (4) MCAO+AT group. Acupuncture procedure was performed for four days. Total RNA was extracted using TRIzol reagent, according to the manufacturer's instructions, and was purified using an RNAeasy mini kit. Immuno-histochemistry was performed using primary antibody mouse anti-BrdU, NeuN, Dcx, and VEGF. Results : We found that $ST_{36}$ had the more neuroprotective effect than $LI_{11}$ and $SP_3$. The microarray analysis revealed that 54 genes were more expressed neurogenesis pathway in MCAO+AT group compared with MCAO group(fold changes greater than or equal to twofold change). 11 genes were more expressed angiogenesis pathway. And 7 genes were more expressed VEGF pathway. Immuno-histochemistry revealed that cell proliferation, cell migration and cell maturation were increased. Conclusions : This study demonstrated that acupuncture on $ST_{36}$ had neuro-protective and neuro-restorative effect in ischemic brain injuries. And its mechanism might be related to promote neurogenesis and angiogenesis. These results suggest that acupuncture have potential benefits for the treatment of ischemic stroke.

Effects of Gastrodiae Rhizoma on Brain Edema and Aquaporin Expressions Following Intracerebral Hemorrhage in Rats (천마(天麻)가 뇌조직출혈(腦組織出血) 흰쥐의 뇌부종(腦浮腫)과 Aquaporins 발현에 미치는 영향)

  • Lee, Ju-Yong;Ku, Ja-Seung;Lee, Dong-Eun;Shin, Jung-Won;Kim, Seung-Joon;Sohn, Nak-Won
    • The Korea Journal of Herbology
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    • v.25 no.4
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    • pp.85-93
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    • 2010
  • Objectives : This study aimed at evaluation of the effects of Gastrodiae Rhizoma on brain edema and aquaporin water channel expressions in the brain. Methods : Brain edema following intracerebral hemorrhage (ICH) was induced by the stereotaxic intrastriatal injection of bacterial collagenase type VII in Sprague-Dawley rats. Then ethanol extract of Gastrodiae rhizoma was treated once a day for 3 days. Brain edema % and water contents, and cell size of neurons in the cerebral cortex were examined. Immuno-histochemistry was processed for AQP4, AQP1, and AQP9 expressions in the brain sections and area % of immuno-labeling was analyzed with image analysis. Results : 1. Ethanol extract of Gastrodiae Rhizoma reduced brain edema of ICH induced rats significantly. 2. Ethanol extract of Gastrodiae Rhizoma reduced excessive brain tissue water contents of ICH induced rats significantly. 3. Ethanol extract of Gastrodiae Rhizoma reduced cellular edema of neurons in cerebral cortex of ICH induced rats significantly. 4. Ethanol extract of Gastrodiae Rhizoma reduced AQP4 immuno-positive area % in cerebral cortex and external capsule of ICH induced rat brain significantly. 5. Ethanol extract of Gastrodiae Rhizoma reduced AQP9 immuno-positive area % in glia limitans externa of ICH induced rat brain significantly. Conclusions : These results suggest that Gastrodiae Rhizoma reveals protective effects against brain edema and cytotoxic edema of neurons by means of down-regulation of AQP4 expression in the brain.

The Effects of Needle Electrode Electrical Stimulation on Cellular Necrosis Blocking the Forebrain after Induction of Ischemia

  • Kim, Sung-Won;Lee, Jung-Sook;Park, Seung-Gyu;Kang, Han-Ju;Kim, Yong-Soo;Yoon, Young-Dae;Yang, Hoe-Song;Lee, Han-Gi;Kim, Sang-Soo
    • Journal of International Academy of Physical Therapy Research
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    • v.1 no.1
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    • pp.10-18
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    • 2010
  • This study was performed to investigate the effects of Needle Electrode Electrical Stimulation(NEES) on ischemia-induced cerebrovascular accidents. After obstruction and reperfusion of arteries in white mice, the amounts of necrosis and inflammation related substances Bax, IL-6, Caspase-3, and COX-2 were measured in neurons of the fore-brain. The following results were obtained. This study used 21 male specific pathogen free(SPF) SD rats, 8 weeks of age and approximately 300g in weight. Each exposed artery was completely occluded with non-absorbent suture thread and kept in that state for 5 minutes. The sutures were then removed to allow reperfusion of blood. Test group is control group(common carotid artery occlusion models), a GI(underwent common carotid artery occlusion), and NEES(underwent NEES after artery occlusion). The GI and NEES groups were given 12, 24, or 48 hours of reperfusion before NEES. NEES device(PG6, ITO, Japan, 9V, current, 2Hz) was used to stimulate the bilateral acupoint ST36 of the SD rats for 30 minutes while they were sedated with 3% isoflurane. An immuno-histochemistry test was done on the forebrains of the GI induced rats. Both Bax and Caspase-3 immuno-reactive cells, related to apoptosis, were greater in the GI than the NEES group. Cox-2 and IL-6 immuno-reactive cells, related to inflammation, were greater in the GI and NEES groups than the control group. We can expect that applying NEES after ischemic CVA is effective for preventing brain cells from being destroyed. And we can conclude NEES should be applyed on early stage of ischemic CVA.

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Immunohistochemical studies in the intestine of the duck, Anas platyrhynchos platyrhyncos Linne, with ages (연령에 따른 청둥오리 장관 내분비세포에 대한 면역조직화학적 연구)

  • Lee, Jae-hyun;Ku, Sae-kwang;Lee, Hyeung-sik
    • Korean Journal of Veterinary Research
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    • v.38 no.1
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    • pp.1-8
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    • 1998
  • The relative frequency and distribution of bovine chromogranin(BCG)-, serotonin-, somatostatin(SOM)- and S-100 protein-immunoreactive cells in the intestinal tract of the duck (Anas platyrhynchos platyrhyncos, Linne) with ages were studied using immunohistochemical methods. BCG-immunoreactive cells were detected in duodenum on 23 days of incubation. Thereafter these cells were occurred throughout the intestine. SOM-immunoreactive cells were detected in the small intestine from hatching to 9 weeks. Thereafter they were also observed in colon. Serotonin-immunoreactive cells were detected throughout the intestinal tracts after hatching. These immunoreactive cells were increased with ages except that BCG-immunoreactive cells were decreased with ages from 6 weeks after hatching. BCG- and serotonin-immunoreactive cells were most frequently detected in colon regions and SOM-immunoreactive cells in duodenum regions, but no S-100 protein-immunoreactive cells were detected in this study.

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Effects of Root of Cibotii Rhizoma on Neuronal Damage of Spinal Cord Contusion Injury in Rats (구척(狗脊)이 흰쥐의 척수압박에 의한 신경세포 손상에 미치는 영향)

  • Park, Won-Sang;Kim, Eun-Seok;Shin, Jung-Won;Kim, Bum-Hoi;Kim, Seong-Joon;Kang, Hee;Sohn, Nak-Won
    • Journal of Korean Medicine Rehabilitation
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    • v.20 no.2
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    • pp.1-15
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    • 2010
  • Objectives : This study was performed to evaluate the effects of root of Cibotii rhizoma(CR) ethanol extract on the tissue and neuronal damage of the spinal cord injury(SCI). Methods : SCI was induced by mechanical contusion following laminectomy of 10th thoracic vertebra in Sprague-Dawley rats. CR was orally given once a day for 7 days after SCI. Tissue damage and nerve fiber degeneration were examined with cresyl violet and luxol fast blue(LFS) histochemistry. HSP72(as neuronal damage marker), MAP2(as nerve fiber degeneration marker), c-Fos(immediate early gene), and Bax(pro-apoptotic molecule) expressions were examined using immuno-histochemistry. Individual immuno-positive cells expressing HSP72, MAP2, c-Fos and Bax were observed on the damaged level and the upper thoracic and lower lumbar spinal segments. Results : 1. CR reduced degeneration of nerve fibers and motor neuron shrinkage in the ventral horn of the lower lumbar spinal segment, but generally it did not seem to ameliorate the tissue injury following SCI. 2. CR reduced demyelination in the ventral and lateral funiculus of the lower lumbar spinal segment. 3. CR reduced HSP72 expression on the neurons in the peri-central canal gray matter adjacent to the damaged region. 4. CR strengthened MAP2 expression on the motor neurons in the ventral horn and on nerve fibers in the lateral funiculus of the lower lumbar spinal segment. 5. CR reduced c-Fos positive cells in the peri-lesion and the dorsal horn of the damaged level and in the ventral horn of the lower lumbar spinal segment. 6. CR reduced Bax positive cells in the peri-lesion and the dorsal horn of the damaged level and in the ventral horn of the lower lumbar spinal segment. Conclusions : These results suggest that CR plays an inhibitory role against secondary neuronal damage and nerve fiber degeneration. following SCI.

Effects of Hwangnyeonhaedok-tang on Cerebral Hemorrhage and Edema in Collagenase Induced-ICH Rats (황련해독탕(黃連解毒湯)이 Collagenase에 의한 흰쥐의 뇌출혈 및 뇌부종에 미치는 영향)

  • Ku, Ja-Seung;Lee, Joon-Suk;Shin, Jung-Won;Kim, Seong-Joon;Sohn, Nak-Won
    • The Journal of Korean Medicine
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    • v.32 no.5
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    • pp.100-113
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    • 2011
  • Objectives: Brain edema is brain swelling that occurs due to the accumulation of excess water in the brain parenchyma. AQP4 and AQP9 are water-channel proteins expressed strongly in the brain and control water fluxes into and out of the brain parenchyma. This study was conducted to evaluate effect of Hwangnyeonhaedok-tang on brain edema and intracerebral hemorrhage. Methods: Intracerebral hemorrhage was induced by intrastriatal injection of type IV collagenase(0.23 U/${\mu}l$, 0.1 ${\mu}l$/min) into Sprague-Dawley rat brains. Hwangnyeonhaedok-tang water extract(1,000 mg/kg) was administered orally three times every 20 hours from 4 hours after ICH operation. Then hematoma volume, brain edema percentage, and water content of brain tissue were measured. Immuno-histochemistry was performed for AQP4 and AQP9 expressions in the brain sections and area % of immuno-labeling was analyzed with image analyzing system. Results: 1. Water extract of Hwangnyeonhaedok-tang reduced hematoma volume of ICH induced rat. 2. Water extract of Hwangnyeonhaedok-tang reduced MPO positive neutrophils in the perihematoma of the ICH induced rat. 3. Water extract of Hwangnyeonhaedok-tang reduced brain edema percentage and water content of brain tissue of ICH induced rat. 4. Water extract of Hwangnyeonhaedok-tang reduced AQP4 immuno-positive cells in the perihematoma of the ICH induced rat. 5. Water extract of Hwangnyeonhaedok-tang reduced AQP9 immuno-positive cells in the perihematoma of the ICH induced rat. Conclusions: These results suggest that Hwangnyeonhaedok-tang decreases intracerebral hemorrhage and brain edema by means of downregulating AQP4 and AQP9 expressions in the brain.

Correlation of Hormone Receptor and HER-2/neu Expression with Clinicopathologic Parameters in Primary Breast Tumors

  • Shaikh, Fouzia;Jamal, Qamar;Baig, Saeeda;Hadi, Naila Irum;Majeed, Noman
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3363-3367
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    • 2016
  • Background: Breast cancer (BC) is a major health issue worldwide as well as in Pakistan. All women belonging to any race, ethnicity or lineage are in danger of developing breast cancer. Significant factors influencing the development of breast malignancies are the genetic background, environmental conditions, reproductive parameters, the consequences of female hormones both intrinsic and extrinsic, alteration of immune status, and biologic determinants. Materials and Methods: Overall 150 biopsy proven patients were included in the study. Samples were submitted for histopathology and determination of estrogen and progesterone receptor expression and HER-2/neu status. Associations with other characteristics like age, tumor stage, node involvement, histological grade were also studied. Results: Mean age at presentation was 46.7 years. The majority had invasive ductal carcinoma, 100 (84.7%), and were in stage pT3, 54 (45.7%). Important relationships (P<0.05) were found among ER, PR positivity, and Her 2 neu overexpression. However, no noteworthy link was identified amongst ER, PR, Her 2 neu and tumor grade, stage, age, lymph node involvement except for the menopausal status. Conclusions: In summary, breast cancer patients featured an advanced stage of disease, more lymph node involvement, and moderately high grade tumors and with more estrogen, progesterone receptor and HER-2 positive tumors.

Effect of Radix Scutellariae on Nicotine Addiction (황금(黃芩)이 니코틴 중독에 미치는 효과)

  • Chang, Gyu-Tae;Kim, Jang-Hyun;Seo, Young-Min
    • The Journal of Pediatrics of Korean Medicine
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    • v.20 no.1
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    • pp.137-149
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    • 2006
  • Objective : The purpose of this study is to investigate the effect of Radix Scutellariae on repeated nicotine-induced locomotor activity and c-Fos expression utilizing Fos-like immuno-histochemistry method in the nucleus accumbens, and the striatum, one of the major projection areas of the control DA system. Methods : Male Sprague-Dawley rats were divided into untreated(normal), nicotine-treated (control), Radix Scutellariae-treated(sample) groups, RS group received Radix Scutellariae(100mg/kg, i.p.) 30minutes before injection of nicotine(0.4mg/kg, s.c.) for 7days. Rat were followed withdrawal for 3 days and one challenge for 1day. Results : Systemic challenge with nicotine produced a much larger locomotor activity and expression of c-Fos in the nucleus accumbens and the striatum. Pretreatment with Radix Scutellariae decreased in nicotine-induced locomotor activity and c-Fos expression in the core, shell, straitum area. Conclusion : These results demonstrated that reduction in locomotor activity by Radix Scutellariae may be mediated by reduction of dopamine release and of postsynaptic neuronal activity in striatum, the nucleus accumbens. Out results show neurochemical evidence for the biological effects of Radix Scutellariae that ultimately may help us to understand how Radix Scutellariae can be used to treat nicotine addiction.

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Effects of Scutellariae Radix on Neuronal Apoptosis of Spinal Cord Contusion Injury in Rats (황금(黃芩)이 척수압박손상 흰쥐의 척수신경세포 자연사에 미치는 영향)

  • Bahn, Hyo-Jung;Jo, Jong-Jin;Kim, Bum-Hoi;Park, Seong-Ha;Shin, Jung-Won;Kim, Seong-Joon;Sohn, Nak-Won
    • Journal of Korean Medicine Rehabilitation
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    • v.21 no.4
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    • pp.13-22
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    • 2011
  • Objectives : This Study was performed to evaluate the effects of Scutellariae Radix(SR) water-extract on the tissue and neuronal apoptosis of the spinal cord injury(SCI). Methods: SCI was induced by mechanical contusion following laminectomy of 10th thoracic vertebra in Sprague-Dawley rats. SR was orally given once a day for 7 days after SCI. Neuronal apoptosis was examined with terminal deoxynucleotidyl transferase-mediated dUTPnick-end labeling(TUNEL) assay. Bax (Bcl-2-asociated X protein), Bcl-2(B-cell blastoma 2), c-Fos(FBJ osteosarcoma oncogene) expressions were examined using immuno-histochemistry. Individual TUNEL and immuno-labeled cells expressing Bax, Bcl-2 and c-Fos were counted on the same level in peri-damaged region and in ventral horn. Results: 1. SR significantly reduced number of TUNEL labeled apoptotic cells induced by the spinal cord contusion injury. 2. SR significantly reduced Bax positive cells expression on the motor neuron in the ventral horn induced by the spinal cord contusion injury. 3. SR strengthened Bcl-2 expression on the motor neuron in the ventral horn induced by the spinal cord contusion injury. 4. SR reduced c-Fos expression on the motor neuron in the ventral horn induced by the spinal cord contusion injury. Conclusions : These results suggest that SR plays an inhibitory role against neuronal apoptosis and has significant effects for locomotor disfunction induced by SCI.

Effect of Yukmijihwangtang on Learning and Memory Impairment in Transient Focal Cerebral Ischemia Rat Model (육미지황탕(六味地黃湯)이 국소뇌허혈유발 기억장애(記憶障碍) 모델 흰쥐에 미치는 영향)

  • Kim, Ki-Hyun;Min, Sang-Yeon;Kim, Jang-Hyun
    • The Journal of Korean Medicine
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    • v.30 no.2
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    • pp.1-16
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    • 2009
  • Objectives: This study investigated the effect of Yukmijihwangtang on cerebral ischemia-induced learning and memory impairment by middle cerebral artery (MCA) occlusion in rats. Methods: The ability of learning and memory of rats was measured using the eight-arm radial maze and the passive avoidance test, and profile of cholinergic neuron was assessed in the medial septum and hippocampus region by immuno-histochemistry. Results: 1. No differences were found between groups in the number of correct choices in acquisition performance during the eight-arm radial maze task. 2. No differences were found between groups on day 1 in the error rate in acquisition performance, which is defined as the number of enters into the same arm more than once within five minutes. After 5 to 6 days of test, the number of errors was significantly reduced in the Yukmijihwangtang group (forebrain ischemia group with Yukmijihwangtang treatment), compared with the ischemia group. 3. The memory processes significantly improved in the Yukmijihwangtang group according to results of the passive avoidance test. 4. The appearance of AchE (acetylcholinesterase) in the CA1 region of hippocampus significantly decreased in the ischemia group, compared with the sham group (untreated group). The appearance of AchE in the same region significantly increased in the Yukmijihwangtang group, compared with the ischemia group. 5. The appearance of ChAT (choline acetyltransferase) in the CA1 region of the hippocampus and medial septum decreased in the ischemia group, compared with the sham group. The appearance of ChAT in the same region significantly increased in the Yukmijihwangtang group, compared with the ischemia group Conclusions: This study provides evidence that Yukmijihwangtang is effective for reviving the ability of learning and memory and damaged neurons in rats with experimental cerebral ischemia.

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