- Anterior Neck Approach for 4-vessel Occlusion in Rats: A Study for Immunohistological Changes and the Effects of Aminoguanidine on Neuronal Cell Death in the Hippocampus.
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Young Jun Ahn, Ki Young Choi, Gu Kang, Yong Seok Kim, Seungkoo Lee, Il Young Cheong, Keun Woo Kim
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Korean J Pathol. 2007;41(6):393-405.
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- BACKGROUND
Clinically relevant cerebral ischemia is encountered most frequently as a cardiac arrest or as single or multiple occlusions of the intracranial or extracranial cerebral arteries. Yamaguchi et al. has introduced a one-stage anterior approach to occlude the common carotid arteries (CCAs) and vertebral arteries (VAs).
METHODS
We used a 2-stage anterior approach for producing transient global ischemia by 4-vessel occlusion (4-VO). Four to five days after electrocauterization of two VAs using the anterior neck approach, two CCAs were clipped for 10 min under anesthesia. Aminoguanidine (100 mg/kg) was administered intraperitoneally immediately after 4-VO, and then twice a day for three consecutive days. Cresyl violet staining and immunohistochemical analysis for the expression of GFAP, CD11b, nitrotyrosine, iNOS, and Bax were performed, using brain slices obtained from the rats that were sacrificed 1, 3, 5 and 7 days after reperfusion.
RESULTS
Aminoguanidine reduced neuronal cell death in the CA1 region of the hippocampus. Expression of GFAP, CD11b, nitrotyrosine, iNOS, and Bax were significantly increased in the CA1 region of the hippocampus three days after 4-VO.
CONCLUSIONS
We believe that modified 4-VO is a good method to study transient forebrain ischemia as it is simple and inexpensive to perform and can be utilized without stereotaxis, a pivoting dissection microscope, EEG, a laser flowmeter or the use of Mongolian gerbils.
- Nicotinamide Reduces the Infarct Volume in a Rat Model of Transient Middle Cerebral Artery Occlusion.
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Min Sup Lee, Young Jun Ahn, Ki Young Choi, Gu Kang, Seong Sik Kang, Il Young Cheong, Kun Jai Lee, Keun Woo Kim
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Korean J Pathol. 2006;40(2):93-102.
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Abstract
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- BACKGROUND
Cerebral ischemia depletes ATP and causes irreversible tissue injury. Nicotinamide is a precursor of NAD+ and it is also a poly (ADP-ribose) polymerase (PARP) inhibitor that increases the neuronal ATP concentration and so protects against stroke. Therefore we examined whether nicotinamide could protect against cerebral ischemia by using a model of transient middle cerebral artery occlusion (MCAO) (reperfusion 2 h post ischemia) in Sprague-Dawley rats.
METHODS
Nicotinamide (500 mg/kg) or normal saline was administered intraperitoneally 24 and 0 h before and after MCAO, respectively. The infarction volumes were determined with triphenyltetrazolium chloride staining 24 h after reperfusion. The nitrotyrosine, PAR polymer and PARP-1 expressions were examined by immunohistochemistry with using brain slices obtained from the rats that were sacrificed at 0, 15, 30, 60 and 120 min after reperfusion.
RESULTS
The infarction volumes were significantly attenuated (21.8%, p<0.05). The nitrotyrosine expressions were increased at 0, 15 and 30 min, and those expressions for PARP polymer and PARP-1 were increased at 60 and 120 min, respectively. Nicotinamide partly reduced the expressions for nitrotyrosine and PAR polymer except for PARP-1.
CONCLUSIONS
These results suggest that nicotinamide may attenuate ischemic brain injury through its antioxidant activity and the inhibition of PARP-1.
- Minocycline Attenuates the Development of Allodynia: An Immunohistochemical Study on CD11b, GFAP and c-Fos in the Spinal Dorsal Horn in SD Rat.
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Gu Kang, Ki Young Choi, Min Sup Lee, Young Jun Ahn, Seong Sik Kang, Il Young Cheong, Wanjoo Chun, Sung Soo Kim
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Korean J Pathol. 2004;38(5):311-318.
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Abstract
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- BACKGROUND
Minocycline, a semisynthetic second-generation tetracycline, is an antibiotic that has excellent ability to penetration into the CNS via the brain-blood barrier.
Minocycline has emerged as a potent inhibitor of microglial activation, and it is an effective neuroprotective agent in experimental brain ischemia. Glial cell activation and proliferation are known to be associated with neuropathic pain in the peripheral nerve injuries.
METHODS
The fifty percent threshold of withdrawal responses was measured in the hindpaws of SD rats following tight ligation of left fifth lumbar spinal nerve. Rats were sacrificed at 1, 3, 5, and 7 days and at 0.5, 1, 2, and 4 h post ligation (n=5/group/time point). Immunohistochemistry for GFAP, CD11b and c-Fos was done on the spinal cord at the level of the fifth lumbar nerve. Minocycline (45 mg/kg) and normal saline (300-400 microL) were administered intraperitoneally, 1 day and 1 h before the operations, and every day postoperatively until the rats were sacrificed.
RESULTS
Treatment with minocycline reduced allodynia and the expressions of CD11b at 5 days and c-Fos at 1 and 2 h post operation compared with the saline treatment (control).
CONCLUSIONS: It was thought that minocycline reduced the allodynia induced by tight ligation of the fifth lumbar spinal nerve in rats through the inhibition of microglial activation and c-Fos expression.
- The Expression of Bcl-2, Bax, Cytochrome C and Caspase-3 in Camptothecin-Induced Apoptosis of Mouse 3T3 Fibroblasts.
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Young Jun Ahn, Min Sup Lee, Gu Kang
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Korean J Pathol. 2002;36(2):71-76.
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Abstract
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- BACKGROUND
Camptothecin (CPT), which has been used for cancer treatment and apoptosis as an inhibitor of DNA topoisomerase I. We investigated the possibility that camptothecin induces anti-appoptotic bcl-2 and pro-apoptotic bax, cytochrome c and caspase-3.
METHODS
We performed immunocytochemical stains for bcl-2, bax and cytochrome c, and also performed westem blots for caspase-3 and the three proteins above using mouse 3T3 fibroblasts treated with CPT (0.5 microgram/mL). The immunostain for bcl-2 was done 12 hours after a microinjection of antisense oligomer to bcl-2 in the nuclei of the cells.
RESULTS
On immunocytochemistry, bcl-2 showed no expressions regardless of CPT treatment and microinjection of the antisense oligomer. The expression of cytochrome c was not changed before and after CPT treatment, and bax demonstrated weak or moderate expressions at 36 and 48 hours afte the treatment. There were no expressions at 0, 12, and 24 hours after CPT treatment. On westem blot, bcl-2 exhibited no expressions before and after CPT treatment. Expressions of ctyochrome c and caspase-3 increased after CPT treatment, and expressions of bax decreased 24 hours after CPT treatment followed by a tendency of increased expressions as time went by.
CONCLUSIONS
In the CPT-induced apoptosis of mouse 3T3 fibroblasts, CPT induced increased expressions of bax, cytochrome c and caspase-3 with no expressions of bcl-2, which are associated with the apoptosis pathway.
- A Study of Apoptosis Induced by Microinjection of Cytochrome c Protein into Mouse 3T3 Fibroblast.
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Min Sup Lee, Gu Kang
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Korean J Pathol. 2002;36(1):1-6.
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Abstract
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- BACKGROUND
Microinjectors have been used for cell biology and development, and are useful for the study of cellular morphologic changes with response to the external milieu and intracellularly injected molecules.
METHODS
This study was performed to confirm the apoptotic changes induced by intracytoplasmic microinjection of cytochrome c (5 mg/mL) to mouse 3T3 fibroblasts with and without pretreatment of Ac-DEVD-CHO (100 mol/mL), and BSA (bovine serum albumin, 5 mg/mL) as a control, and evaluate the usefulness of microinjection as a method to study apoptosis pathways.
RESULTS
Mild focal cytoplasmic fragmentation was seen in the cells microinjected with cytochrome c as early as 10 min after the injection. Apoptotic morphology with apoptotic body formation was observed at 60 min after the injection, and then new apoptotic change of the injected cells was not seen. Cytochrome c-injected cells showed about 31% of apoptotic cells of the total injected cells 50-60 min after the injection. BSA-injected cells did not show apoptosis morphology at 50-60 min after the injections. Caspase-3 inhibitor, Ac-DEVD-CHO-treated cells with cytochrome c microinjection exhibited lower apoptosis indices (average apoptosis index; 11.5+/-8.6%) than non-treated cells of the inhibitor (average apoptosis index; 11.5+/-8.6%).
CONCLUSIONS
It was observed that intracellular microinjection of cytochromic c induced apoptosis which was inhibited by Ac-DEVD-CHO, although apoptotic cells were so easily detached that further study could not be performed.
However it is thought that microinjection should be a method to study apoptosis and signal transduction with the molecular biological techniques currently available.
- Correlation of Tumor Angiogenesis and nm23-H1 Expression with Lymph Node Metastasis in Proper Muscle Gastric Cancer.
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Eun Sook Nam, Gu Kang, Hyung Sik Shin, Young Eui Park
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Korean J Pathol. 1997;31(5):410-416.
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- We studied clinicopathologic features of 44 cases of PM (proper muscle) gastric cancer, correlated the lymph node metastasis and found the result of immunohistochemical staining for tumor angiogenesis using antibodies to Factor VIII-related antigen and nm23-H1, known as meatastasis inhibitory substance. The results were as follows: 1) The average age of these 44 cases of PM gastric cancer was 55.1 years old (range 35-81). The ratio of male to female was 2.2 : 1. The tumor was located at the antrum of stomach in 72.7% of the cases. The average size of the tumor was 4.1 cm (range 0.6-9). The gross features were comprised of Borrmann type I (6.8%), II (29.6%), III (56.8%), IV (6.8%), respectively. The microscopic type was a diffuse type in 70.5% and an intestinal type in 29.5%. There were lymph node metastasis in 25 of the 44 cases (56.8%). 2) The microvessel count was higher in the lymph node positive group (average 69.3) than in the lymph node negative group (average 45.6) (P=0.004). There was a higher microvessel density in diffuse type, over 4 cm of tumor size, proximally located tumor, older than 50 years, Borrmann type II and IV, but there was no statistically significant correlation. 3) The more decreased expression of nm23-H1 was found in the lymph node positive group (56.0%) than in the lymph node negative group (31.6%), but showed no statistical significance (P=0.0142).
There was no significant correlation between the expression of nm23-H1 and the other clinicopathologic factors. We suggest that the microvessel count of the tumor angiogenesis may be a prognostic factor for predicting lymph node metastasis and also help to determine the therapeutic modalities of PM gastric cancer.
- Cytologic Findings of Epithelial-Myoepithelial Carcinoma of the Salivary Gland: A Case Report.
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Eun Sook Nam, Gu Kang, Hyung Sik Shin
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Korean J Cytopathol. 1996;7(1):64-68.
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- The report of aspiration cytologic findings of epithelial-myoepithelial carcinoma(EMC) in the salivary gland is extremely rare. We present a case of fine needle aspiration cytology(FNAC) from EMC in the right submandibular gland of a 46 years old male patient. Neck CT scan revealed a confined lesion in the submandibular gland without enlargement of the regional lymph node. FNAC from the tumor showed several three-dimensional cellular clusters with admixed normal acinar cells. They frequently formed branching tubular structures composed of two type of cells; darker cells having eosinophilic scanty cytoplasm with round dense nuclei and clear cells having abudant pale cytoplasm with vesicular nuclei at the periphery of clusters. The tumor cells of both types did not show pleomorphism or mitoses. The resected submandibular gland showed an ill-defined whitish firm tumor, measuring 2 X1.5X2cm. The histology revealed an infiltrative tumor showing characteristic two cell types in a duct-like arrangement surrounded by thin basement menbrane. An inner layer of darker cells and outer layer of clear cells were postive for cytokeratin in the former and S-100 protein in the latter on the immunohistochemical stain.
- Immunohistochemical Analysis of Transforming Growth Factor-beta Expression in Gastric Adenocarcinoma.
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Young Hee Choi, Seoung Wan Chae, Min Chul Lee, Jung Weon Shim, Hye Kyung Ahn, Hye Rim Park, Gu Kang, Hyung Sik Shin, Young Euy Park
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Korean J Pathol. 1994;28(3):272-281.
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Abstract
- Thirty cases of gastric adenocarcinoma were examined immunohistochemically for expression of transforming growth factor-beta(TGF-beta) in order to analyze significant correlation with clinical stage and pathologic grade of gastric adenocarcinoma. Specific immunostaining was clearly detected in the cytoplasm of the neoplastic cells. The TGF-,6 expression in the gastric adenocarcinoma is closely related to the depth of invasion, the degree of invasiveness and the presence of metastasis. Thus, we observed the stronger immunohistochemical. expression of TGF-beta in the deeper portion of invasion and in the invasive gastric adenocarcinomas with the lymph nodal metastasis than in the superficial portion of invasion and in those without the lymph nodal metastasis. There results suggest that the transforming growth factors expression in carcinoma cells may play an important role in the carcinomatous invasion resulting in metastasis.
- Pathologic Classification of the Resected Gastric Carcinoma.
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Gu Kang, Hyung Sik Shin, Min Chul Lee, Young Euy Park, Joo Seop Kim, Chul Jae Park, Soo Tong Pai
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Korean J Pathol. 1992;26(1):17-27.
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- A total of 212 cases of gastric carcinoma resected at Kang-Dong Sacred Heart Hospital during the period of 4 years from December 1986 to December 1990 were examined according to Borrmann, Mulligan-Rember, Ming and WHO methods based on histopathological investigations. In Mulligan-Rember (M-R) classification, intestinal cell type (IC) was frequently seen in Borrmann type I and II; pyloro-cardiac gland cell type (PC) in II and III, and mucous cell type (MC) in IV and III. Expanding growth pattern was more frequent in IC, infiltrative growth pattern in MC, and PC showed less infiltrative growth than MC. In gross type, the most expanding growth pattern was seen in Advanced gastric carcinoma type (AGC) I and the next one was in AGC II; the ratio of infiltrative versus expanding type was highest in AGC IV and next in AGC resembling early gastric carcinoma (EGC) and AGC III in order. On WHO classification except squamous type, all the papillary type showed expanding growth and infiltrative growth was frequently seen in signet-ring cell, undifferentiated, tubular and mucinous types in order. Lymphoid stroma was more frequently found in expanding type than infiltrative type. The frequency of angioinvasion of tumor cells observed was high in AGC resembling EGC, AGC II, III, IV, I and EGC in order. In WHO classification excluding squamous type, undifferentiated and signet-ring cell types occurred more frequently under the age of 60 and papillary type were more frequnetly seen over the age of 60. But tubular type had no difference between the two age groups. In Ming's classification, expanding type was more frequently seen than infiltrative type over the age of 60.
- Two Patterns of Gastric Carcinoma with Lymphoid Stroma.
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Gu Kang, Eun Sil Yu, Yong Il Kim
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Korean J Pathol. 1988;22(4):375-382.
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- Various clinical and pathologic parameters in order to determine the prognosis of gastric carcinoma have been proposed. Among them "carcinoma with lymphoid stroma" has been proven to show good prognosis. But the criteria of lymphoid stroma in this condition remain vague and not clear. A total of 7 cases of gastric carcinoma with heavy lymphoid stromal response out of 947 surgically resected gastric carcinomas was reviwed with histotopographic analysis. They were all advanced carcinoma, Borrmann type I and II. Histologically, the lymphoid stromal response could be divided into three patterns; nodular (3 cases), diffuse (3 cases) and mixed (1 case). The nodular pattern was characterized by massive lymphoid cell infiltration with many follicle formation and little desmoplastic reaction, while the diffuse pattern showed diffuse permeative type of inflammatory cell infiltration with scarce lymphoid follicle formation and mild desmoplasia. Regional lymph node metastasis was found in 2 cases; one in diffused and another one in mixed pattern. The stromal reaction was not directly related with the depth of tumor invasion. We propoose that the term GCLS should be used in the cases of nodular pattern with complete follicle formation of lymphoid stroma.
- Subglottic Laryngeal Atresia: Associated with Tracheoesophageal Fistula.
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Gu Kang, Je G Chi
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Korean J Pathol. 1988;22(2):145-147.
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- A case of congenital laryngeal atresia of infraglottic type in a newborn infant is described for is rarity and importance of early diagnosis. This case was associated with tracheoesophageal fistula, and died in 5 minutes of life due to asphxia. Failure to intubate in the neonate should raise the possibility of laryngeal atresia and emergency management should be provided. The lung tissue was normally developing despite the complete obstruction of the outlet of the air and possible amniotic fluid passage.
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