• Title/Summary/Keyword: immunohistochemical methods

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The Effect of SuJeom-san on Caerulein-induced Acute Pancreatitis in Rats (Caerulein으로 유발된 흰쥐의 급성 췌장염에 대한 수념산(手拈散)의 효과)

  • Park, Jae-Seok;Choi, Chang-Won
    • The Journal of Internal Korean Medicine
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    • v.31 no.3
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    • pp.500-512
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    • 2010
  • Objectives : This study was designed to investigate the effects of SuJeom-san(SJS) extract in rats with caerulein-induced acute pancreatitis (AP). Methods : We examined changes of pancreatic weight, histological, immunohistochemical and gene expression of cyclooxygenase (COX-2). Thirty-six adult male Sprague-Dawley rats were divided into six groups as follow: normal(Nor), caerulein-induced (Con), caerulein + cefotaxime sodium(CT), caerulein + SJS 3 mg/kg(SJSA), caerulein + SJS 6 mg/kg(SJSB) and caerulein + SJS 12 mg/kg(SJSC) groups. Pancreatic tissues of rats from all groups were removed for histological observation and light, and electron microscopic examination. Platelet activating factor(PAF) and Interleukin-6(IL-6) levels were determined spectrophotometrically. Results : The ratio of pancreas/body weight was significantly(p<0.05) increased in the Con compared with Nor, but significantly(p<0.05) decreased in SJSA, SJSB, SJSC and CT groups compared with Con. Caerulein administration significantly increased(p<0.05) the levels of amylase, but SJSA, SJSB, SJSC and CT significantly(p<0.05) reduced the levels of these enzymes. The levels of platelet activating factor(PAF) increased in Con compared with Nor, but decreased in SJSA, SJSB, SJSC and CT groups compared with Con. Interleukin-6(IL-6) levels increased significantly in all groups compared to Nor at 6 hrs, but significantly(p<0.05) reduced in SJSA, SJSB, SJSC and CT groups compared with Con at 24 hrs. The levels of tumor necrosis factor(TNF)-${\alpha}$ levels increased in all groups compared to Nor at 6 hrs, but significantly(p<0.05) reduced in SJSA, SJSB, SJSC and CT groups compared with Con at 24 hrs. The COX-2 positive materials were observed in the pancreas of the Con, but these positive materials were decreased in the SJS extract treatment group. Conclusion : SJS is potentially capable of limiting pancreatic damage during AP by restoring the fine structure of acinar cells and tissue; therefore, we conclude that SJS may have beneficial effects in the treatment of caerulein-induced AP.

Allopregnanolone suppresses mechanical allodynia and internalization of neurokinin-1 receptors at the spinal dorsal horn in a rat postoperative pain model

  • Fujita, Masahide;Fukuda, Taeko;Sato, Yasuhiro;Takasusuki, Toshifumi;Tanaka, Makoto
    • The Korean Journal of Pain
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    • v.31 no.1
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    • pp.10-15
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    • 2018
  • Background: To identify a new strategy for postoperative pain management, we investigated the analgesic effects of allopregnanolone (Allo) in an incisional pain model, and also assessed its effects on the activities of the primary afferent fibers at the dorsal horn. Methods: In experiment 1, 45 rats were assigned to Control, Allo small-dose (0.16 mg/kg), and Allo large-dose (1.6 mg/kg) groups (n = 15 in each). The weight bearing and mechanical withdrawal thresholds of the hind limb were measured before and at 2, 24, 48, and 168 h after Brennan's surgery. In experiment 2, 16 rats were assigned to Control and Allo (0.16 mg/kg) groups (n = 8 in each). The degree of spontaneous pain was measured using the grimace scale after the surgery. Activities of the primary afferent fibers in the spinal cord (L6) were evaluated using immunohistochemical staining. Results: In experiment 1, the withdrawal threshold of the Allo small-dose group was significantly higher than that of the Control group at 2 h after surgery. Intergroup differences in weight bearing were not significant. In experiment 2, intergroup differences in the grimace scale scores were not significant. Substance P release in the Allo (0.16 mg/kg) group was significantly lower than that in the Control group. Conclusions: Systemic administration of Allo inhibited mechanical allodynia and activities of the primary afferent fibers at the dorsal horn in a rat postoperative pain model. Allo was proposed as a candidate for postoperative pain management.

Effects of Rehmanniae Radix on HPA Axis System and Catecholamienrgic System in FST (생지황(生地黃)이 강제수영부하시험에서 HPA Axis System과 Catecholaminergic System에 미치는 영향)

  • Park, Jae-Beom;Lee, Tae-Hee
    • Herbal Formula Science
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    • v.18 no.1
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    • pp.133-144
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    • 2010
  • Object : The effects of Rehmanniae Radix(RR) on HPA Axis system and Catecholaminergic system was investigated. Methods : we performed the Forced Swimming Test(FST). Also the expression of Corticotropin -Releasing Factor(CRF), c-Fos and Tyrosine Hydorxylase(TH) was measured by immunohistochemical method at Paraventricular Nucleus(PVN), Locus Coeruleus(LC) and Ventral Tegmental Area(VTA). Results : 1. The duration of immobility in the Forced Swimming Test was significantly decreased in the Rehmanniae Radix 400 mg/kg groups in comparison with the control group(p<0.05). 2. Comparing to the control group, CRF expression was significantly decreased in the Rehmanniae Radix 400 mg/kg treated group in comparison with the control group(p<0.01). 3. c-Fos expression was significantly decreased at PVN in the Rehmanniae Radix 100 mg/kg treated group in comparison with the control group(p<0.01). 4. TH expression was significantly increased at LC in the Rehmanniae Radix 100 mg/kg and 400 mg/kg treated group in comparison with the control group(p<0.001). 5. TH expression was significantly decreased at VTA in the Rehmanniae Radix 100 mg/kg and 400 mg/kg treated group in comparison with the control group(p<0.001). Conclusion : According to the results, it can be considered that Rehmanniae Radix has antidepressant effect by showing the reduction of immobility in FST through the decreased expression of CRF, c-Fos in PVN and TH in VTA.

The Immunologic Expression of Apoptosis Related Proteins and Apoptotic Cells in Human Ovarian Follicles (인간난포에서 Apoptosis 관련 단백질 및 Apoptotic 세포의 면역학적 발현)

  • Park, Sung-Rok;Lee, Byung-Seok;Yang, Woo-Ick;Kim, Jong-Hwa;Park, Byung-Joo;Park, Ki-Hyun;Cho, Dong-Jae;Song, Chan-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.3
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    • pp.195-200
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    • 2002
  • Objective : To investigate the expression of apoptosis related proteins and apoptotic cells on the human ovarian follicles. Materials and Methods: Thirty five Formalin-fixed paraffin-embedded human ovarian tissue blocks were selected from the surgical pathology files of the department of pathology, College of Medicine, Yonsei University, for the period from 1996 to 1998. All specimen were from premenopausal women aged from $32{\sim}45$. Ovarian tissues were collected from the patients performing hysterectomy for benign uterine diseases. Immunohistochemical staining was performed for the detection of DNA fragmented cell, Bcl-2, Bax, Fas and Fas-ligand. Results: Bcl-2 and bax were not expressed on the surrounding cells and oocyte of the primary, primordial and preantral follicles. Fas and Fas-ligand (Fas-L) were not expressed on the surrounding cells on the primordial and primary follicles. But expressed on the surrounding granulosa cells and oocyte in the primordial and primary follicles. In the healthy follicles, Bcl-2 was expressed on the granulosa cells, however, Bax was not expressed. DNA fragmented cells were expressed on the inner granulosa cell layer of atretic follicles. Conclusion: Fas, Fas-ligand, and Bax may be responsible for the follicular atresia and Bcl-2 may be involved in the follicular survival in the human ovary.

COX-2 Expression in Renal Cell Carcinoma and Correlations with Tumor Grade, Stage and Patient Prognosis

  • Tabriz, Hedieh Moradi;Mirzaalizadeh, Marzieh;Gooran, Shahram;Niki, Farzaneh;Jabri, Maryam
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.535-538
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    • 2016
  • Background: Cyclooxygenase 2 (COX-2) is important as an enzyme in the pathway leading to the production of prostaglandin E2 (PGE2) and arachidonic acid. This pathway is known to play a role in inflammation, tumor growth, invasiveness and metastasis, inhibition of apoptosis and angiogenesis. Inhibition of COX-2 has been shown to be a promising antitumor and antiangiogenic strategy in several tumor types, including renal cell carcinoma (RCC). Therefore, we decided to evaluate the immunohistochemical expression of this marker and its association with several clinicopathological characteristics in a series of cases. Materials and Methods: COX-2 expression was examined immunohistochemically in tumor tissues obtained from 96 patients who underwent radical (94 cases) or partial (2 cases) nephrectomy. Correlations between COX-2 expression and clinicopathologic findings including pathologic stage, nuclear grade and other indicator of prognosis were examined. Results: Of 96 tumors, 20.9% were positive for COX-2 expression. A correlation was found between COX-2 expression and tumor histological subtype (P=0.03).The papillary subtype showed maximum expression of this marker (43.8%) and the clear subtype minimum (14.7%). There were also possible links between COX-2 expression and pathologic stage, nuclear grade and nodal involvement but the results were not statistically significant (P=0.8, P= 0.14 and P=0.06, respectively). No correlation was found between COX2 expression and patient age, gender, tumor size, metastasis or survival. Conclusions: In our study, COX-2 expression was correlated with the histological subtype of RCC. Additional research is required to determine the link between COX-2 expression and prognosis and also evaluation of probable effectiveness of COX-2 inhibitor drugs in treatment of RCC patients.

The Polymerase Chain Reaction in Diagnosis of Small B-Cell Non-Hodgkin Lymphomas

  • Antoro, Ester Lianawati;Dwianingsih, Ery Kus;Indrawati, Indrawati;Triningsih, FX Ediati;Harijadi, Harijadi
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.491-495
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    • 2016
  • Background: Small B-cell non-Hodgkins lymphoma (NHL) is difficult to be distinguished from non-neoplastic reactive processes using conventional haematoxylin-eosin (HE) staining due to different interpretations among pathologists with diagnosis based on morphologic features. Ancillary examinations such as immunohistochemical (IHC) staining are essential. However, negative or doubtful results are still sometimes obtained due to unsatisfactory tissue processing or IHC technique. The polymerase chain reaction (PCR) as a molecular diagnostic technique is very sensitive and specific. Clonality detection of heavy chain immunoglobulin (IgH) gene rearrangement has been widely used to establish diagnosis of B-cell NHL. Aims: To elaborate interobserver variation in small B-cell NHL diagnosis based on morphologic features only and to confirm sensitivity and specificity of the PCR technique as an ancillary method. Materials and Methods: A toptal of 28 samples of small B cell NHL and suspicious lymphoma were interpreted by 3 pathologists in Sardjito General Hospital based on their morphology only. The reliability of assessment and the coefficient of interobserver agreement were calculated by Fleiss kappa statistics. Interpretation results were confirmed with IHC staining (CD20, CD3, Bcl2). PCR was performed to analyze the clonality of IgH gene rearrangement. Results: Interobserver agreement in morphologic evalution of small B cell NHL and chronic lymphadenitis revealed kappa coefficient 0.69 included in the substantial agreement category. The cases were divided into 3 groups based on morphology and IHC results; lymphoma, reactive process and undetermined group. PCR analysis showed 90% sensitivity and 60% specificity. Conclusions: The present study revealed a substantial agreement among pathologists in small B-cell NHL diagnosis. For difficult cases, PCR is useful as complementary method to morphologic and IHC examinations to establish definitive diagnosis.

Study on the Correlation between Extracellular Matrix Protein-1 and the Growth, Metastasis and Angiogenesis of Laryngeal Carcinoma

  • Meng, Xin-Yu;Liu, Juan;Lv, Feng;Liu, Ming-Qiu;Wan, Jing-Ming
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2313-2316
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    • 2015
  • Objective: To investigate the correlation between extracellular matrix protein-1 (ECM1) and the growth, metastasis and angiogenesis of laryngeal carcinoma. Materials and Methods: Forty-five samples with laryngeal benign and malignant tumors confirmed by pathology in Laiwu City People's Hospital from March 2006 to March 2011 were collected, in which there were 29 cases with laryngeal carcinoma and 16 with benign tumors. The expression of ECM1 and factor VIII-related antigens in patients with laryngeal carcinoma and those with benign tumors was respectively detected using immunohistochemical method, and the correlation between ECM1 staining grade and microvessel density (MVD) was analyzed. Results: In laryngeal carcinoma tissue, ECM1 was mainly expressed in cytoplasm, less in cytomembrane or intercellular substance. With abundant expression in the tissue of laryngeal benign tumors (benign mesenchymoma and hemangioma), ECM1 was primarily expressed in the connective tissue, which was different from the expression in laryngeal carcinoma tissue. The proportion of positive ECM1 staining (++) in patients with laryngeal carcinoma was dramatically higher than those with benign tumors (p<0.05), and that of strongly-positive ECM1 staining (+++) slightly higher. The results of Spearman nonparametric correlation analysis revealed that ECM1 staining grade in laryngeal carcinoma tissue had a significantly-positive correlation with MVD (r=0.866, p=0.000). Conclusions: ECM1 expression in laryngeal carcinoma is closely associated with tumor cell growth, metastasis and angiogenesis, which can be considered as an effective predictor in the occurrence and postoperative recurrence of laryngeal carcinoma.

Diagnostic and Clinical Significance of KIT(CD117) Expression in Thymic Epithelial Tumors in China

  • Song, Nan;Chen, Gang;Zhang, Peng;Liu, Ming;He, Wen-Xin;Jiang, Ge-Ning
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2745-2748
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    • 2012
  • Aims: To study KIT (CD117) expression in thymic epithelial tumors in China, and investigate diagnostic and clinical significance. Material and Methods: Thymic epithelial tumors (TETs) from 102 patients (3 type A, 29 type AB, 5 type B1, 22 type B2, 29 typeB3 and 16 thymic carcinomas) were examined. Immunohistochemical staining with an antic-kit monoclonal antibody was performed on a tissue microarray. Relationships between KIT positive expression and the TET clinical characteristics (WHO histologic classification and Masaoka stage system) were analysed. Results: The KIT positive expression rate was significantly higher in thymic carcinoma (60%, 9/16) than in thymoma (8%, 7/86), a strong correlation being found with the WHO classification, but not the Masaoka tumor stage. The overall survival for patients with KIT positive lesions was significantly worse. Conclusions: KIT is a good molecule marker to differentially diagnose thymic carcinoma from thymoma, while also serving as a predictor of prognosis for TETs. Further research into KIT mutations in Chinese TETs should be conducted to assess the efficacy of targeted therapy.

Relationship Between PTEN and Livin Expression and Malignancy of Renal Cell Carcinomas

  • Cheng, Tao;Zhang, Jian-Guo;Cheng, Yuan-He;Gao, Zhong-Wei;Ren, Xiao-Qiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2681-2685
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    • 2012
  • Objectives: This study focused on PTEN and Livin expression and associations with malignancy in human renal clear cell carcinomas (RCCC). Methods: PTEN and Livin expression was assessed in 100 RCCC tissue samples, 50 paracarcinoma cases, and 20 normal renal tissue samples using the immunohistochemical Streptavidin proxidase (SP) method. The relationships between binding and corresponding biological characteristics, such as histological grade, lymph node metastases, and clinical stages were analyzed. Results: Positive PTEN expression in RCCC was significantly lower than in renal tissue adjacent to carcinoma tissue and normal renal tissue (P<0.01). Livin expression in the renal tissue adjacent to the carcinoma and normal renal tissues exhibited only low levels, whereas overall Livin expression in RCCC was statistically significant (P<0.01). In RCCC, PTEN expression rate gradually decreased with an increase in clinical stage, whereas that of Livin increased to statistically significant levels (P<0.01), PTEN and Livin levels being negatively correlated (r=-0.395, P<0.01). Conclusions: PTEN and Livin are important in RCCC development. The two factors combined are expected to provide indices for estimating RCCC malignancy and progression levels, as well as references for RCCC diagnosis and treatment.

Ovarian Sex Cord Stromal Tumours in Children and Young Girls - A More Than Two Decade Clinicopathological Experience in a Developing Country, Pakistan

  • Haroon, Saroona;Idrees, Romana;Zia, Aleena;Memon, Aisha;Fatima, Saira;Kayani, Naila
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1351-1355
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    • 2014
  • Background: Ovarian sex-cord stromal tumours (SCST) are rare, and relatively infrequent in children. These have to be distinguished from more common germ cell tumors in children and also from benign epithelial neoplasms. Objectives: The purpose of our study was to report the clinical and pathological findings in young patients with these tumours in our population. Material and Methods: The present observational cross-sectional study included all subjects <21 years of age diagnosed with ovarian SCST, in Aga Khan University Hospital Histopathology Laboratory, Karachi, Pakistan, from January 1992 till July 2013. Results: Of the total of 513 SCSTs presented during the study period, 39 fulfilled inclusion criteria and were assessed. The age range was 4-250 months. Most of the tumours presented at stage-1 and an abdominal mass was the most common presenting symptom, along with menstrual disturbance. The left side ovary was slightly more affected (53.5%). Of the total, 15 were juvenile granulosa cell tumours (JGCT), 11 sclerosing stromal tumours (SST), 10 of the fibrothecomas spectrum, 2 Sertoli leydig cell tumours (SLCT) and one a sex cord tumour with annular tubules (SCTAT). Detailed immunohistochemical analyses were performed in 33 cases. Recurrence/metastasis was noted in 4/21 cases with follow-up data. Conclusions: Ovarian sex cord stromal tumours are very rare in young age in our population, and usually present at an early stage. Most common among these are juvenile granulosa cell tumours, although surprisingly sclerosing stromal tumours were also common. Clinical symptoms due to hormone secretion in premenstrual girls and menstrual disturbance in menstruating girls are common presenting features.