• Title/Summary/Keyword: histological method

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Gross, organoleptic and histologic assessment of cadaveric equine heads preserved using chemical methods for veterinary surgical teaching

  • Rodrigo Romero Correa;Rubens Peres Mendes;Diego Darley Velasquez Pineros;Aymara Eduarda De Lima;Andre Luis do Valle De Zoppa;Luis Claudio Lopes Correia da Silva;Ricardo de Francisco Strefezzi;Silvio Henrique de Freitas
    • Journal of Veterinary Science
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    • v.25 no.2
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    • pp.29.1-29.11
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    • 2024
  • Background: Preservation of biological tissues has been used since ancient times. Regardless of the method employed, tissue preservation is thought to be a vital step in veterinary surgery teaching and learning. Objectives: This study was designed to determine the usability of chemically preserved cadaveric equine heads for surgical teaching in veterinary medicine. Methods: Six cadaveric equine heads were collected immediately after death or euthanasia and frozen until fixation. Fixation was achieved by using a hypertonic solution consisting of sodium chloride, sodium nitrite and sodium nitrate, and an alcoholic solution containing ethanol and glycerin. Chemically preserved specimens were stored at low temperatures (2℃ to 6℃) in a conventional refrigerator. The specimens were submitted to gross and organoleptic assessment right after fixative solution injection (D0) and within 10, 20, and 30 days of fixation (D10, D20, and D30, respectively). Samples of tissue from skin, tongue, oral vestibule, and masseter muscle were collected for histological evaluation at the same time points. Results: Physical and organoleptic assessments revealed excellent specimen quality (mean scores higher than 4 on a 5-point scale) in most cases. In some specimens, lower scores (3) were assigned to the range of mouth opening, particularly on D0 and D10. A reduced the range of mouth opening may be a limiting factor in teaching activities involving structures located in the oral cavity. Conclusions: The excellent physical, histologic, and organoleptic characteristics of the specimens in this sample support their usability in teaching within the time frame considered. Appropriate physical and organoleptic characteristics (color, texture, odor, and flexibility) of the specimens in this study support the use of the method described for preparation of reusable anatomical specimens.

Study on the Anti-cancer, Anti-metastasis and Immune response improvement Effects of Aqua-acupuncture with Rubi Fructus infusion solution (항암(抗癌) 및 면역효과(免疫效果)에 복분자약침(覆盆子藥鍼)이 미치는 영향(影響))

  • Lee, Seon-Hee;Lee, Byung-Ryul
    • Journal of Acupuncture Research
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    • v.17 no.1
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    • pp.189-219
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    • 2000
  • To study the effects of anti-cancer, anti-metastasis and immune response improvement effects of aqua-acupuncture with Rubi Fructus infusion solution, we used Rubi Fructus infusion solution(taken by water-alcohol method) put into Chung-wan (CV12) and Chok-Samni(ST36) of BALB/c or C57BL/6 which are corresponding to humanbody. We observed the cytotoxicity, the effect on the expression of MMP-9 gene, the ability to control cancer cell proliferation, change of body weight, surviving number, median surviving time, increase of life span, changes in amount of leukocyte, erythrocyte, platelet, total protein, creatinine, glucose and LDH, weight of spleen, number of pulmonary colony, histological analysis on tissue metastasis of lung and liver, splenic cell proliferation, the expression of cytokine gene, the number of $CD4^+$, $CD8^+$, $CD19^+$ and NK cell, and concluded like this. The results were obtained as follows : 1. Effects of Anti-cancer 1) The cytotoxicity about B16-F10 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-5}$, $2^{-6}$, $2^{-7}$, $2^{-8}$ diluent groups in Rubi Fructus infusion solution treatment was inhibited significantly, compared with control group. 2) The cytotoxicity about HT1080 cell line of $2^0{\sim}2^{-8}$ diluent groups in Rubi Fructus infusion solution treatment was inhibited significantly, compared with control group. 3) The effect on expression of MMP-9 gene was inhibited significantly in all the sample groups, compared with control group. 4) The effect on the control-ability on the cancer cell proliferation showed cytotoxicity significantly in $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-4}$, $2^{-5}$, $2^{-6}$, $2^{-7}$, diluent groups. 2. Effects of Anti-metastasis 1) S-180 cancer cell line transplants in BALB/c mice were inhibited significantly in weight increase in all the sample groups, compared with control group. The surviving number increased in almost sample groups, except one group put into Chok-Samni(ST36) with 20% Rubi Fructus infusion solution treatment group that showed same number of the control group. 2) S-180 cancer cell line transplants in BALB/c mice showed high MST significantly in almost sample groups, compared with control group. But one group put into Chok-Samni(ST36) with 20% Rubi Fructus infusion solution showed low MST than control group. 3) The group injected in vein with B16-F10 cancer cell line in C57BL/6 mice showed increased ILS than control group significantly in anti-metastasis test. 3. Effects of Immune response improvement 1) The group injected in vein with B16-F10 cancer cell line in C57BL/6 mice were increased significantly in the number of leukocyte and glucose, and decreased significantly in the amount of platelet and LDH, compared with control group. However, there's no significant increase or decrease in number of erythrocyte, total protein and creatinine. 2) We couldn't find any significant relation in spleen weight of the sample group. 3) In pulmonary colony, sample group was decreased significantly, compared with control group. 4) Histological analysis of sample group inhivited compared with that of control group in both of lung and liver. 5) In immune system, all the sample groups showed having more relevancy to the effect on splenic cell proliferation than normal group. 6) Cytokine gene increased in almost sample groups, except one group treated with $50{\mu}g/m{\ell}$ Rubi Fructus infusion solution on IL-12. 7) In flow cytometry there's no significant relation in number of $CD8^+$ cell, however, the number of $CD4^+$, $CD19^+$ cell and NK cell in sample group had more relation than in control group. Above the results showed that aqua-acupuncture of Rubi Fructus solution has effects of anti-cancer, and-metastasis and immune response improvement.

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Study on the Anti-cancer, Anti-metastasis and Immune response improvement of Aqua-acupuncture with Cistanches Herba infusion solution (육종용약침의 항암작용(抗癌作用) 및 면역효과(免疫效果)에 대(對)한 실험적(實驗的) 연구(硏究))

  • Lee, Eun-Yong;Lee, Byung-Ryul
    • Journal of Acupuncture Research
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    • v.17 no.1
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    • pp.251-286
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    • 2000
  • To study the effects of anti-cancer, anti-metastasis and immune response improvement of aqua-acupuncture with Cistanches Herba infusion solution, we used Cistanches Herba infusion solution(taken by water-alcohol method) put into Chung-wan(CV12) and Chok-Samni(S36) of BALB/c or C57BL/6 which are corresponding to human body. We observed the cytotoxicity, the effect on the expression of MMP-9 gene, the ability to control cancer cell proliferation, change of body weight, surviving number, MST, ILS, changes in amount of WBC, RBC, PLT, total protein, creatinine, glucose and LDH, weight of spleen, number of pulmonary colony, histological analysis on tissue metastasis of lung and liver, splenic cell proliferation, the expression of cytokine gene, the number of $CD4^+$, $CD8^+$, $CD19^+$ and NK cell, and concluded like this. The results were obtained as follows : 1. The cytotoxicity about B16-F10 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-5}$ diluent groups in Cistanches Herba infusion solution treatment was inhibited significantly, compared with control group. 2. The cytotoxicity about HT1080 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-5}$, $2^{-8}$ diluent groups in Cistanches Herba infusion solution treatment was inhibited significantly, compared with control group. 3. The effect on expression of MMP-9 gene was decreased in all the sample groups, compared with control group. 4. The effect on the control-ability on the cancer cell proliferation showed cytotooicity significantly in $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-4}$, $2^{-5}$ diluent groups. 5. S-180 cancer cell line transplants in BALB/c mice were inhibited significantly in weight increase in all the sample groups, compared with control group. The surviving number increased in almost sample groups, except one group put into Chok-Samni(S36) with 20% Cistanches Herba infusion solution treatment group that showed same number of the control group. 6. S-180 cancer cell line transplants in BALB/c mice showed high MST and ILS significantly in almost sample groups, compared with control group. But one group put into Chok-Samni(S36) with 20% Cistanches Herba infusion solution treatment group showed low MST and ILS than control group. 7. The sample group injected in vein with B16-F10 cancer cell line in C57BL/6 mice showed increased ILS compared with control group significantly in anti-metastasis test. 8. The sample group injected in vein with B16-F10 cancer cell line in C57BL/6 mice were increased significantly in the number of WBC and glucose, and decreased significantly in the amount of LDH, compared with control group. However, there's no significant increase or decrease in number of RBC, PLT, total protein and creatinine. 9. We couldn't find any significant relation in spleen weight of the sample group. 10. In pulmonary colony, sample group was decreased significantly, compared with control group. 11. Histological analysis of sample group inhivited compared with that of control group in both of lung and liver. 12. In immune system, all the sample groups showed having more relevancy to the effect on splenic cell proliferation than normal group. 13. The effect on cytokine gene expression of all the sample groups were increased than control group. 14. In flow cytometry there's no significant relation in number of $CD8^+$, $CD19^+$ cell, however, the number of $CD4^+$ cell and NK cell in sample groups were increased than in control group. Above the results showed that aqua-acupuncture of Cistanches Herba infusion solution has effects of anti-cancer, anti-metastasis and immune response improvement.

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The Clinicopathological Factors That Determine a Local Recurrence of Rectal Cancers That Have Been Treated with Surgery and Chemoradiotherapy (직장암의 수술 후 방사선 치료 시 국소 재발의 임상 병리적 예후 인자)

  • Choi, Chul-Won;Kim, Min-Suk;Lee, Seung-Sook;Yoo, Seong-Yul;Cho, Chul-Koo;Yang, Kwang-Mo;Yoo, Hyung-Jun;Seo, Young-Seok;Hwang, Dae-Yong;Moon, Sun-Mi;Kim, Mi-Sook
    • Radiation Oncology Journal
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    • v.24 no.4
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    • pp.255-262
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    • 2006
  • $\underline{Purpose}$: To evaluate the pathological prognostic factors related to local recurrence after radical surgery and adjuvant radiation therapy in advanced rectal cancer. $\underline{Materials\;and\;Methods}$: Fifty-four patients with advanced rectal cancer who were treated with radical surgery followed by adjuvant radiotherapy and chemotherapy between February 1993 and December 2001 were enrolled in this study. Among these patients, 14 patients experienced local recurrence. Tissue specimens of the patients were obtained to determine pathologic parameters such as histological grade, depth of invasion, venous invasion, lymphatic invasion, neural invasion and immunohistopathological analysis for expression of p53, Ki-67, c-erb, ezrin, c-met, phosphorylated S6 kinase, S100A4, and HIF-1 alpha. The correlation of these parameters with the tumor response to radiotherapy was statistically analyzed using the chi-square test, multivariate analysis, and the hierarchical clustering method. $\underline{Results}$: In univariate analysis, the histological tumor grade, venous invasion, invasion depth of the tumor and the over expression of c-met and HIF-1 alpha were accompanied with radioresistance that was found to be statistically significant. In multivariate analysis, venous invasion, invasion depth of tumor and over expression of c-met were also accompanied with radioresistance that was found to be statistically significant. By analysis with hierarchical clustering, the invasion depth of the tumor, and the over expression of c-met and HIF-1 alpha were factors found to be related to local recurrence. Whereas 71.4% of patients with local recurrence had 2 or more these factors, only 27.5% of patients without local recurrence had 2 or more of these factors. $\underline{Conclusion}$: In advanced rectal cancer patients treated by radical surgery and adjuvant chemo-radiation therapy, the poor prognostic factors found to be related to local recurrence were HIF-1 alpha positive, c-met positive, and an invasion depth more than 5.5 mm. A prospective study is necessary to confirm whether these factors would be useful clinical parameters to measure and predict a radio-resistance group of patients.

Comparison of Outcomes after Curative Resection of Primary Lung Cancer between 50 Year or Younger and 70 Year or Older Patients (50세 이하와 70세 이상 원발성 폐암 환자에서의 근치적 수술 후 성적 비교)

  • Lee, Jae-Ik;Kim, Keun-Woo;Park, Kook-Yang;Park, Chul-Hyun;Jeon, Yang-Bin;Choi, Chang-Hyu
    • Journal of Chest Surgery
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    • v.42 no.2
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    • pp.206-213
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    • 2009
  • Background: Previous series have suggested that younger patients with primary lung cancer exhibit a more aggressive disease course with a worse prognosis, as compared to older patients, although this issue is still debatable. Material and Method: We reviewed the medical records of 79 patients (32 patients 50 years and younger (Group I) and 47 patients 70 years and older (Group II)) who underwent curative resection for primary lung cancer between July 2000 and June 2008. Result: The median age of the patients was 46.5 years in Group I and this was 73 years in Group II. The older patients were more likely to have major comorbidities (44% versus 77%, respectively; p=0.003). Histological examinations identified that the minor histological types (excluding non-small cell lung cancer (NSCLC)) were predominantly found in the Group I patients (16% versus 2%, respectively; p=0.037). For the TNM staging of the NSCLC, with excluding the minor histologic types, a higher proportion of patients had stage III disease in Group I (33% versus 13%, respectively; p=0.038). There was no significant difference in major morbidity (16% versus 30%, respectively; p=0.148) and operative mortality (0% versus 4.3%; p=0.512) between the groups. The mean follow-up interval was 33 months (range: $1{\sim}98$ months) for patients in both groups. For the patients with NSCLC, the five-year overall survival rate was 52.3% for Group I and 53.7% for Group II (p=0.955). The rate of freedom from recurrence at five years was significantly lower for the Group I patients than for the Group II patients (39.4% versus 70.4%, respectively; p=0.027), and only being a member of Group I impacted recurrence, based on the Cox proportional hazard analysis (p=0.034). Of the patients who had recurrence, four patients in Group I underwent aggressive surgical treatment. All of these patients exhibited long-term survival (range: $46{\sim}87$ months). Conclusion: In our study, the early outcome and long-term survival were similar for the younger and older patients after curative resection of primary lung cancer. However, we think that younger patients require meticulous follow-up as they had a tendency to proceed to surgery with advanced stage disease, a higher recurrence rate than did the older patients and the survival rates were improved, even for the recurred cases, with early aggressive treatment.

The Inflammatory Response in Mouse Lung after Acute Sulfur Dioxide Exposure (급성 아황산가스 폭로후 흰쥐의 폐에 유발된 염증반응에 관한 연구)

  • Chin, Young-Joo;Park, Nam-Gyu;Lee, Hyeon-Suk;Kim, Dae-Soo;Earm, Jae-Ho;Cho, Myeong-Chan;Yoon, Sei-Jin;Jeong, Hwa-Sook;Song, Hyung-Geun;Sung, Ro-Hyun;Lee, Sang-Do
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.4
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    • pp.328-338
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    • 1994
  • Background: Effect of sulfur dioxide($SO_2$) exposure on airway is well known but little about the effect of $SO_2$ exposure on lung parenchyme. This study is to determine if short tenn exposure to $SO_2$ in concentration commonly found in industrialized environment cause potentially harmful effect on the lung parenchyme, and to evaluate the exposure time-response relationship between short tenn exposure to $SO_2$ and the inflammatory response in mouse lung. Method: 5ppm $SO_2$ gas was used and 48 mice were grouped into control(10), 30(9), 60(11), and 120 minute exposure(18) group. In each group, bronchoalveolar lavage(BAL) was done immediately after and at 1,2,3 days after exposure. Histological examination was performed in control and 120 minute exposure group. Results: 1) Cell response in bronchoalveolar lavage fluid. In 30 and 60 minute exposure group, compared to the control group, lymphocyte count has significantly increased(p<0.05) at 1 day after exposure but did not differ at 2 days after exposure. In 120 minute exposure group, also compared to the control group, there was significant increase in total cell, macrophage, and lymphocyte count at 1 day after exposure, (p<0.05) which lasted for 2 days but did not significantly differ at 3 days after exposure. 2) Histological findings in 120 minute exposure group. In the airway, mild epithelial cell damage and ciliary loss were noted but there was no evidence of inflammatory cell infiltration. Interstitial inflammatory infiltration was noted at 1 day after exposure, which lasted for 3 days after exposure and there was no evidence of edema or fibrosis in the interstitium Conclusions: These data indicate potentially noxious effect of $SO_2$ on the lung parenchyme as well as the airway at exposure level that are regarded as relatively safe, and the duration of injury depends on the exposure time.

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Evaluation of p16INK4a/Ki-67 Dual Immunostaining in Liquid-based Cytology for Diagnosis of Uterine Cervical Dysplasia and Cancer (자궁경부 이형성증과 암의 진단을 위한 액상세포 검체에서 p16INK4a/Ki-67 이중면역염색의 평가)

  • Sung, Mi Hee;Lee, Hoon Taek;Shin, Min Shik;Oh, Seo Young;Kim, Wook Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.3
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    • pp.132-139
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    • 2015
  • Recently, $p16^{INK4a}$/Ki-67 dual immunostaining has been introduced as a new biomarker protocol for early detection of uterine cervical dysplasia and cancer in liquid-based cytology (LBC). We performed the $p16^{INK4a}$/Ki-67 dual immunostaining using a CINtec$^{(R)}$ PLUS kit in a total of 109 LBC cases of cervicovaginal smear and compared its results with those from LBC, HPV hybrid capture II (HC II) test and histological diagnosis. Expression of $p16^{INK4a}$ and Ki-67 was significantly associated with cases of LSIL or higher in cytological diagnosis and cases of cervical intraepithelial neoplasia (CIN) 1 or higher in histological diagnosis (p<0.001 and p<0.001, respectively). Among forty-six cases of atypical squamous cells of undetermined significance (ASCUS) in LBC, $p16^{INK4a}$ and Ki-67 was expressed in 31 (67.4%), which were positively associated with cases of CIN I lesion or higher in histology. The sensitivity of $p16^{INK4a}$/Ki-67 dual immunostaining for finding lesions of CIN 1 or higher was 89.0%, which was higher than LBC. The specificity was 73.5%, which was higher than that of the HC II test. Based on these results, the $p16^{INK4a}$/Ki-67 dual immunostaining method can be a useful diagnostic marker for improving the sensitivity of LBC and the specificity of HC II test.

Usefulness of Tuberculin Skin Test by Tuberculin PPD RT23 2 TU (Tuberculin PPD RT23 2 TU를 이용한 투베르쿨린 피부반응 검사의 의의)

  • Yang, Jong-Wuk;Jeon, Man-Jo;Kim, Sung-Jung;Lee, Hyang-Lim;Lee, Seung-Jun;Lee, Myoung-Goo;Kim, Dong-Gyu;Park, Myoung-Jae;Kang, Min-Jong;Hyoen, In-Gyu;Jung, Ki-Suck
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.4
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    • pp.401-408
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    • 2002
  • Background : Tuberculin skin test is a method to examine M. tuberculosis infection and has been used all over the world. But various factors make it difficult to understand testing results. In 2000, the American Thoracic Society recommended that skin test results should be decided by considering risk factors of the tested. In Korea, high tuberculosis infection rate and BCG vaccination rate make it difficult to differentiate current infection, past infection, and no infection by the skin test. This study was attempted to examine a negative predictive value of the skin test to understand how the skin test acts on deciding administration of anti-tuberculosis drug. Methods : From Mar. 1 to Jul. 31 in 2001, the test was performed for patients hospitalized in Department of Internal Medicine, Hallym University College of Medicine, Chunchon, Korea by administering Tuberculin PPD RT23 2 TU (0.1 ml)to them that has been currently used in Korea based on Mantoux method. They were decided to be infected with tuberculosis bacilli by following diagnostic standard: 1) tuberculosis bacilli was cultured in sputum by microbiological diagnostic standard or Acid-fast bacilli was proven on a microscopic examination or 2) tuberculosis bacilli was not proven in the aforesaid microbiological test by clinical diagnostic standard, while there was opinion or symptom suitable for tuberculosis by radiographic or histological standard so the doctor decided to apply the tuberculosis treatment. Results : In this study, total 210 patients except 20 patients (8.7%) among 230 hospitalized patients were evaluated. Their average age was 60±16.8 years, and male-female rate was 1.28 : 1 (male: 118, female: 92). Number of patient, who was diagnosed and decided as tuberculosis, was 53(25.2%). Pulmonary tuberculosis was found in 45 patients (84.9%); 22 patients were decided to be positive in the Acid-fast bacilli smear test by microbiological examination (culture positive: 13, culture negative: 9), and 23 patients were decided to be tuberculosis patients by clinical diagnosis standard. Tuberculosis pleuritis was found in 8 patients (15.1%); 4 patients were diagnosed and decided by histological standard, and 4 patients were decided and treated by clinical standard. In differentiating patients into 'Negative' and 'Positive' by the skin test standard of the American Thoracic Society, negative predictive value 92.3%, positive predictive value 47.3%, sensitivity and specificity were 83%, 68.8%, respectively. Conclusion : In hospitalized respiratory patients, there was high negative predictive vlaue 92.3% by tuberculin skin test, therefore skin test would be a important factor for deciding administration of anti-tuberculosis drug on negative skin test patient.

Comparative Histological Study on the Parafollicular Cells of Mammals (각종 포유동물 갑상선내의 소포방세포에 관한 비교조직학적 연구)

  • 고정식;박상윤
    • The Korean Journal of Zoology
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    • v.23 no.2
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    • pp.89-108
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    • 1980
  • Comparative studies on the parafollicular cells of the some mammalian species from five different orders were carried out; i.e., man from Primates, cattle, pig, and black goat from Artiodactyla, dog from Carnivora, rabbit from Lagomorpha, rat, mouse, and squirrel from Rodentia. For this study, various special techniques for the parafollicular cells, including Grimelius' silver impregnation method (Sawicki and Bajko, 1974), Singh's argentaffin method (Singh, 1964), HCl-toluidine blue stain (Sawicki, 1971), and HCl-lead hematoxylin stain (Solcia et al., 1969), were applied. Authors obtained the following results: 1. Number of parafollicular cells in the same area of thyroid tissues are significantly different from species to species. Number of cells were largest in dog and less cells were found in the following orders; rat, squirrel, mouse, rabbit, cattle, pig, black goat and finally the smallest number in man. 2. Distribution of parafollicular cells within thyroid gland are significantly different from portion to portion in case of cattle, rabbit, squirrel and mouse, but it is not significant in dog, man, pig, black goat and rat (see Table 1-1 and 1-2). 3. In dog, clustered parafollicular cells are located usually in the interfollicular space, and groups of parafollicular cells are located in the para-and/or inter-follicular positions in rabbit. But in the other animals parafollicular cells are found solitarily in the intra-and/or para-follicular positions. 4. The shape of parafollicular cells shows oval to round contour in dog, but it is polymorphic, for example, spindle, conical, oval, round or elongated with cytoplasmic processes, in the other animals. 5. Size of parafollicular cells is larger in cattle, dog and pig, smaller in rat, mouse and squirrel, and medium-size in rabbit, man and black goat. 6. Parafollicular cells of pig, cattle, dog and squirrel are observed to contain densely packed granules, whereas those of mouse, rat and man contain relatively scanty granules. 7. Parafollicular cells of all the mammals show more or less positive reaction to Grimelius' argyrophile silver impregnation method, HCl-toluidine blue stain and HCl-lead hematoxylin stain, whereas they show negative reaction to argentaffin method (see Table 2). 8. Considering the above finding, it is concluded that there are species differences in the distribution, location and shape of parafollicular cells, and infer that preferable staining method should be selected for reliable detection of parafollicular cells, beacuse staining methods applied on the cells in this study show variable reactions according to species.

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Organ Induction by Combined Dose of bFGF and HGF in Animal Cap Assay of Early Xenopus laevis Embryos. (Xenopus laevis 초기 배의 동물극 분리배양에서 bFGF와 HGF 혼합처리에 의한 기관유도)

  • 진정효;윤춘식;이호선;박용욱;정선우
    • Journal of Life Science
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    • v.14 no.3
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    • pp.375-384
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    • 2004
  • Fibroblast growth factors (FGFs) are known to induce multiple functions in early development, including mesoderm formation, gastrulation movement and antero-posterior patterning. The induction of mesoderm from Xenopus presumptive ectoderm and the combination effect on inducing organs of bFGF(basic FGF) and HGF (Hepatocyte Growth Factor) were studied. Explants were cultured in the combined solution for 3 days to normal embryo arrive at St. 43. These effects on combined dose were examined by histological experiment and by immunohistochemical method. The concentrations of growth factors were tested in 0, 0.5, 1, 10 and also tested in 50 ng/ml of bFGF, and 0, 1, 10, 50 and 100ng/ml of HGF respectively. The synergistic effects were seen in the combined-dose of bFGF and HGF rather than in single dose. Various organs were differentiated and highest inducing effects were seen at the combined concentration of 1 ng/ml of bFGF and 10ng/ml of HGF, and at the concentration 10ng/ml of bFGF and 1 ng/ml of HGF. The bFGF induces various organs from cultured animal cap explants and the effects are time and dose-dependent. HGF is also a potent mitogen for renal tubular cells and for mature hepatocytes in primary culture. Eyes were developed in high percentage at the combined concentration of 1 and 10ng/ml of bFGF, and 1 and 10 ng/ml of HGF. From the induced eye and normal embryonic eye, RPE65 was commonly detected by monoclonal antibodies 40All and 25F5 and the localization of RPE65 was seen by AP reaction.