• Cement
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• s.57
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• pp.16-18
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• 1974

• Journal of Microbiology and Biotechnology
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• v.17 no.12
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• pp.1955-1964
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• 2007

A recent report showed that analysis of CD154 expression in the presence of the secretion inhibitor Brefeldin A (Bref A) could be used to assess the entire repertoire of antigen-specific $CD4^+\;T$ helper cells. However, the capacity of intracellular CD154 expression to identify antigen-specific $CD8^+\;T$ cells has yet to be investigated. In this study, we compared the ability of intracellular CD154 expression to assess antigen-specific $CD8^+\;T$ cells with that of accepted standard assays, namely intracellular cytokine IFN-${\gamma}$ staining (ICS) and MHC class I tetramer staining. The detection of intracellular CD154 molecules in the presence of Bref A reflected the kinetic trend of antigen-specific $CD8^+\;T$ cell number, but unfortunately showed less sensitivity than ICS and tetramer staining. However, ICS levels peaked and saturated 8 h after antigenic stimulation in the presence of Bref A and then declined, whereas intracellular CD154 expression peaked by 8 h and maintained the saturated level up to 24 h post-stimulation. Moreover, intracellular CD154 expression in antigen-specific $CD8^+\;T$ cells developed in the absence of $CD4^+\;T$ cells changed little, whereas the number of IFN-${\gamma}$-producing $CD8^+\;T$ cells decreased abruptly. These results suggest that intracellular CD154 could aid the assessment of antigen-specific $CD8^+\;T$ cells, but does not have as much ability to identify heterogeneous $CD4^+\;T$ helper cells. Therefore, the combined analytical techniques of ICS and tetramer staining together with intracellular CD154 assays may be able to provide useful information on the accurate phenotype and functionality of antigen-specific $CD8^+\;T$ cells.

• Journal of the Korean Arthroscopy Society
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• v.9 no.2
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• pp.154-161
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• 2005

Purpose: to describe the histologic appearance of the type III bone bruise in knees which had sustained an acute anterior cruciate ligament (ACL) rupture. Materials and Method: Twenty-five patients who sustained acute ACL rupture were prospectively enrolled in this study. On MRI, 14 patients demonstrated type III bone bruise on lateral femoral condyle, and 11 patients didn't demonstrated bone bruise. Arthroscopic evaluation and biopsy of the articular cartilage and subchondral bone wert performed before ACL reconstruction. Histologic and immunohistochemical evaluations were done. Results: There was no difference between the bone bruise and control group in the hematoxylin-eosin staining for cell distribution, Masson's trichrome staining for collagen and immunohistochemical staining for type I and type II collagen (p>0.05). But in the safranin-O staining for glycosaminoglycan distribution, the bone bruise group had an evidence of decreased staining at the superficial and middle layers, compared with the control group (p<0.05). We also found fatty change of bone marrow in calcified zone of the bone bruise group with safranin-O staining. Conclusion: We suggest that the type III bone bruise found on MRI indicates a substantial damage to normal articular cartilage homeostasis, and may induce further damage of the articular cartilage.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.4
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• pp.371-377
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• 2016

Decalcification is routinely performed to obtain a pathological diagnosis using bone marrow biopsy. During the decalcification process using a conventional acidic solution, such as HCl, the antigenicity of tissue is damaged. Especially DNA and RNA in the bone marrow are impaired. Hence, there is the need for a standardized decalcification protocol that preserves the antigenicity of tissue. To this end, we compared the effects of two commonly used decalcifiers: Commercial decalcifier (Calcl-Clear Rapid, HCl) and the EDTA (12.5%, pH 7.0). Bone marrow biopsies sampled from 71 patients were decalcified in accordance with the protocols of respective groups-HCI versus EDTA. The differences of decalcification protocols were analyzed with respect to Hematoxylin & Eosin staining, Gomori'sreticulum staining, and immunohistochemical staining and molecular analysis. Immunohistochemical staining used Ki-67, CD20 and CD138 as primary antibodies and molecular analysis was conducted through the DNA concentration analysis, in situ hybridization (ISH) and immunoglobulin heavy chain (IGH) gene rearrangement. On the routine histopathology analysis, there was no difference between HCl and EDTA. Moreover, in case of immunohistochemical staining, the cytoplasmic membrane or cytoplasmic CD markers was well preserved. However, nuclear proteins, such as Ki-67, were stained with low quality. Conversely, according to the molecular analysis, the EDTA protocol preserved the DNA and RNA compared with the HCI. The differences of DNA quantity and quality were statistically significant between protocols of HCl and EDTA. We used 38 cases in HCl and 12 cases in EDTA. Consequently, the EDTA protocol maintains the antigenicity of the protein on tissue and is acceptable for examination with molecular base analysis. Decalcification of bone marrow biopsy by EDTA is highly recommended for the examination of immunohistochemical staining and molecular analysis.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.2
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• pp.551-557
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• 2015

Background: Differentiating morphologic features based on hematoxylin-eosin (HE) staining is the most common method to classify pathological subtypes of non-small-cell lung cancer (NSCLC). However, its accuracy and inter-observer reproducibility in pathological diagnosis of poorly differentiated NSCLC remained to be improved. Materials and Methods: We attempted to explore the role of immunohistochemistry (IHC) staining in diagnosing pulmonary squamous cell carcinoma (SQCC) with poorly differentiated features by HE staining or with elevated serum adenocarcinoma-specific tumor markers (AD-TMs). We also compared the difference of epidermal growth factor receptor (EGFR) mutation rate between patients with confirmed SQCC and those with revised pathological subtype. Logistic regression analyses were used to test the association between different factors and diagnostic accuracy. Results: A total of 132 patients who met the eligible criteria and had adequate specimens for IHC confirmation were included. Pathological revised cases in poor differentiated subgroup, biopsy samples and high-level AD-TMs cases were more than those with high/moderate differentiation, surgical specimens and normal-level AD-TMs. Moreover, biopsy sample was a significant factor decreasing diagnostic accuracy of pathological subtype (OR, 4.037; 95% CI 1.446-11.267, p=0.008). Additionally, EGFR mutation rate was higher in patients with pathological diagnostic changes than those with confirmed SQCC (16.7% vs 4.4%, p=0.157). Conclusions: Diagnosis based on HE staining only might cause pathological misinterpretation in NSCLC patients with poor differentiation or high-level AD-TMs, especially those with biopsy samples. HE staining and IHC should be combined as pathological diagnostic standard. The occurrence of EGFR mutations in pulmonary SQCC might be overestimated.

• Journal of Radiation Protection and Research
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• v.29 no.1
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• pp.9-15
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• 2004

Firstly, we compared the two staining techniques, Giemsa and Acridine orange, to determine micronuclei on samples of cultures of five healthy human peripheral blood lymphocytes after ${\gamma}-irradiation\;(^{137}Cs)$ in dose ranges of 0 to 800cGy. It was found that the Acridine orange staining method gives more reliable results than the usual Giemsa staining method in micronucleus tests. Moreover, the frequency of micronuclei in cytokinesis-blocked human B-lymphocytes was studied after in vitro irradiation in dose ranges of 0 to 50cGy. After setting and separating the B-lymphocytes, the frequency of radiation-induced micronuclei were observed as the end-point markers for the low-dose radiation dosimetry after staining with Giemsa and Acridine orange dyes. The micronuclei frequency in B-lymphocytes was significantly elevated from 10 to 30cGy ${\gamma}-irradiation$. The determination of micronuclei in B-lymphocytes after staining with Acridine orange was higher than that of Giemsa. The frequency of micronuclei in B-lymphocytes was observed to be at least two times higher than those of T-lymphocytes Giemsa in dose increasing. Therefore, the determination of low-dose radiation-induced micronuclei in B-lymphocytes after staining with Acridine orange is likely to have the greatest potential in the estimation of low dose radiation exposure.

• The Journal of Korean Academy of Prosthodontics
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• v.33 no.4
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• pp.611-633
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• 1995

The pupose of this study was to compare the fracture strength of five kinds of all-ceramic crowns(Vintage, Dicor Empress-staining, Empress-layering, In-Ceram) luted with glass ionomer cerment and composite resin cement and to evaluate the effect of cements on the fracture stregth of all ceramic crowns. Five groups of twelve uniform sized all-ceramic crown specimens were fabricated. Six specimens of each group were cemented with glass ionomer cement(Fuji G.I. Cement) and the remaining six specimens of each group were etched, silane-treated, and cemented with composite resin cement(Bistite resin cement). The crowns were stored in water$(37^{\circ}C)$ for 1 day prior to loading in an Instron, using a steel ball(diameter 4mm) at a crosshead speed of 0.5mm/min. The crowns were angled $30^{\circ}$, so the steel ball contacted with the crowns 2mm lingual from the mid-incisal edge. The results obtained were summarized as follows ; 1. With G.I. cement, mean fracture load(Kg) Were : Intage : $18.33{\pm}1.47$ ; Empress-staining : $23.92{\pm}6.67$ ; Dicor : $24.0{\pm}5.81$ ; Empress-layering : $26.92{\pm}2.80$ ; In-Ceram : $51.58{\pm}6.87$ ; ANOVA revealed a significant difference existed(p<0.05) between the group A(Vintage, Dicor, Empress-staining, Empress-layering) and group B(In-Ceram). 2. With Resin cement, mean fracture load(Kg) were : Intage : $22.75{\pm}4.97$ ; Dicor : $42.75{\pm}7.07$ ; Empress-staining : $44.08{\pm}7.99$ ; Empresslayering : $50.42{\pm}5.43$ ; In-Ceram : $52.58{\pm}6.51$ ; ANOVA revealed a significatnt difference existed(p<0.05) between the group A(Vintage) and B(Dicor, Empress-staining Empress-alyering, In-Ceram). 3. Resin cement significantly increased the fracture strength of the all-ceramic crowns for Dicor(156%), Empress-staining(185%), Empress-alyering(187%)(p<0.05); but did not increase the fracture strength of Vintage(128%) and In-Ceram(101%)(p>0.05). 4. Majority of the all-ceramic crowns show a wedge fracture extending through proximal surfaces to an apex, usually apical third(with G.I. cement) or middle third(with Resin cement) of the facial surface.

• Applied Microscopy
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• v.31 no.1
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• pp.1-8
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• 2001

The morphological features of neuronal dendritic spines are changed their shapes, sizes and density in response to physiological or pathological conditions . Therefore, exact analysis of spines warrants understanding of neuronal function. The size of the spine is at the borderline of resolution with light microscopy. High voltage electron microscopy Provide excellent resolution of the spines with proper stain techniques thanks to its higher resolution and penetration power. We evaluated more effective staining method for observing dendritic spines after labeling Purkinje cells with anti-calbindin 28 kD immunohistochemistry or Golgi staining methods. 4 fm thickness sections were observed with high voltage electron microscopy and some morphometric analyses were performed. Both Golgi staining and immunohistochemistry revealed the detail structures of the Purkinje cell such as soma, dendrites, and dendritic spines. High voltage electron micrographs with Golgi staining provide more precise morphology and are easy to measure. Average density of spine is $24.5{\pm}3.6/10{\mu}m$ and its length is $1.12{\pm}0.22{\mu}m$. For quantitative analysis of the spines, high voltage electron, micrographs with Golgi staining are more effective. This preliminary result is expected to be useful for further study of spine plasticity in various conditions.

• Tuberculosis and Respiratory Diseases
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• v.48 no.3
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• pp.324-330
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• 2000

Objective : Lung cancer arises after a series of morphological changes, which take several years to progress from normal epithelium to invasive cancer. Multiple molecular changes and growth factor production have been documented in lung cancers, both small cell and non-small cell types. Insulin-like growth factors(IGFs) are important mitogenic and anabolic peptides, both in vivo and in vitro, and are thought to be significant autocrine-paracrine factors involved in normal and malignant cell proliferation. In this study, the degree of expression of IGF-1 on the immunohistochemical staining in human non-small cell lung cancer(NSCLC) cells and small cell lung cancer (SCLC) cells were investigated. Methods : Immunohistochemical staining for IGF-1 was performed in 15 cases of small cell carcinoma, 15 cases of squamous cell carcinoma, 15 cases of adenocarcinoma, and 12 cases of bronchoalveolar carcinoma. Results : The expression of IGF-1 on the immunohistochemical staining significantly increased in NSCLC cells than in SCLC cells. Conclusion : These results suggest the expression of IGF-1 in human lung cancer cells. The immunohistochemical staining of IGF-1 in lung cancer cell lines may assist in the differentiation of NSCLC and SCLC.

• Journal of Oral Medicine and Pain
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• v.3 no.1
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• pp.23-27
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• 1977

The author thought that the results of cytologic tests could be changed according to the fixative methods. So the author has studied several methods of fixing exfolaiative cells collected from normal adult cheek mucosa with a solution of equal parts of ether and 95% ethylalcohol, 95 ethylalcohol and Isoprophyl alcohol, and compared the results with each other. The results were as follows : The effects of fixation, and staining efficiency of nucleus and protoplasm are similar in each method, but the staining efficiency of cytoplasm and the conversation of cytoplasmic membrane were best in 95% ethylalcohol.