• 제목/요약/키워드: PCNA

검색결과 223건 처리시간 0.034초

한우 섬유아세포의 성 판별 및 세포주기 유도 분석 (Sexing and Cell Cycle Induction Hanwoo Fetal Fibroblast Cells)

  • 김현주;강회성;최화식;이성호;박창식;진동일
    • 한국가축번식학회지
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    • 제27권1호
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    • pp.53-59
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    • 2003
  • 본 연구에서는 한우 태아의 시기별로 35일령, 50일령, 70일령 및 90일령의 fetal fibroblast cell line을 생산하였고, bovine-specific primer와 Y chromosome-specific primer를 이용하여 PCR에 의해 성을 판별하여 각각 암수 2 line의 한우 fetal fibroblast cell line을 확립하였다. 이들 cell line을 계대배양하여 passage number가 10 이상에서 염색체 분석을 실시하였는데 모두에서 80%이상의 세포가 60개의 정상 염색체수의 나타내어 계대배양이 karyotype에 영향을 미치지 않는 것으로 나타났다. Serum starvation과 confluent 배양 방법을 이용하여 Go 상태로 유도되었는지 확인하기 위해 PCNA antibody를 이용하여 Western blotting 분석을 실시하였는데 PCNA 발현이 현저히 감소되는 것을 확인할 수 있었고, 다시 정상 medium으로 환원시켰을 때 세포분열이 재개되어 Go상태로 유도되었음을 확인할 수 있었다. 또한 serum stravation 방법이 conflent한 배양방법보다 PCNA 발현양이 적은 것으로 나타나 좀더 효율적인 Go 상태 세포 주기 조절방법으로 판명되었다.

Transcriptional Regulation of the Drosophila Proliferating Cell Nuclear Antigen Gene and raf Proto-oncogene by Ursolic Acid in Drosophila Cultured Kc Cells

  • Park, Thae-Yeong;Rhee, Sang-Hoon;Kim, Han-Do;Kim, Chong-Rak;Kang, Ho-Sung;Yoo, Mi-Ae
    • Animal cells and systems
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    • 제1권1호
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    • pp.151-155
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    • 1997
  • Promoter of the Drosophila proliferating cell nuclear antigen (PCNA) gene contains DRE (Drosophila DNA replication-related element) required for the high level expression of replication-related genes. Recently, we found that promoter region of the D-raf (a Drosophila homolog of the human c-raf-1) contains two sequences homologous to the DRE and demonstrated the DRE/DREF (DRE-binding factor) involvement in regulation of the D-raf gene. In this study, using ursolic acid (UA), a pentacyclic triterpene acid reported to possess antitumor activities, we examined effects of UA on proliferation of the Drosophila cultured Kc cells and on expression of the PCNA and D-raf genes. UA showed an inhibitory effect on proliferation of the Kc cells in a concentration-dependent manner in DNA content assays and [3H]thymidine incorporation assays. The IC50 value of anti-proliferative effects of UA in DNA content assays was about 7.5uM. UA showed inhibitory effects on expression of the PCNA as well as on that of the D-raf, which were examined with the reporter plasmic p5'-168DPCNACAT or p5'-878DrafCAT, respectively. The results obtained in the present study suggest that expression of the PCNA and D-raf genes is coordinately regulated in at least UA-treated Kc cells and that down-regulation of expression of the PCNA and D-raf genes might be related with the antitumor activities of UA.

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Ginsenosides Promote Proliferation of Cultured Ovarian Germ Cells Involving Protein Kinase C-mediated System in Embryonic Chickens

  • Liu, Hongyun;Zhang, Caiqiao
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권7호
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    • pp.958-963
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    • 2006
  • The effect of ginsenosides (GS) on germ cell proliferation was evaluated with a chicken ovarian germ-somatic cell coculture model and the mechanism involving protein kinase C (PKC) pathway was investigated. Ovarian cells were cultured in serum-free McCoy's 5A medium and challenged with GS alone or in combinations with PKC activator (phorbol 12-myristate 13-acetate, PMA) or inhibitor ($H_7$) for 48 h. The number of germ cells was counted and the proliferating cells were identified by immunocytochemistry of proliferating cell nuclear antigen (PCNA). Results showed that GS significantly increased germ cell proliferation and this stimulating effect was further increased by PMA, but inhibited by H7, in a dose-dependent manner. Moreover, GS-elevated PCNA expression and the PCNA -labeling index of germ cells displayed similar changes with the increased numbers of germ cells. These results indicated that GS stimulated proliferation of ovarian germ cells with involvement of the PKC-mediated system.

항암제 cyclophosphamide가 diethylnitrosamine에 의한 랫드 간암에 미치는 조직학적 영향 (Histological effect of cyclophosphamide on diethylnitrosamine-induced hepatic tumors in rats)

  • 곽수동;강정부;고필옥;김종섭
    • 대한수의학회지
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    • 제39권3호
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    • pp.593-601
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    • 1999
  • This study was designed to evaluate the effect of cyclophosphamide(CY) on diethylnitrosamine(DEN)-induced hepatic tumors in rats. Thirty five male or female Sprague Dawley rats were continiously given water containing 0.01% DEN for 10 weeks and then were given with CY 25mg/rat/day in water for 3, 5, 7 or 9 days. The livers of rats were removed and fixed in 10% buffered neutral formalin. he appearances of positive cells by immunohistochemical methods using proliferating cell nuclear antigen (PCNA) antibody, p53 antibody and apoptotic kit were investigated. The livers of rats given with CY were grossly brilliant, red-brown color, flexible, and thin border, and stainability of the liver cells were restored microscopically, and the vaccuolated and degenerated regions were differentiated from restored regions. These restored findings also were advanced in control group because of no DEN treatment but tended to be less avanced. In immunohistochemistry, positive cells to PCNA antibody appeared more numerous in control groups than that of CY treated groups. Appearance of positive cells in CY-treated group for 7 days and for 9 days were more numerous than those of CY-treated groups for 3 days and for 5 days, respectively. So these findings suggested that CY suppressed cell proliferations and effects of these action were decreased with CY-treated days. The numbers of positive cells to PCNA antibody were more prominent in hepatocellular carcinoma regions and cholangiocarcinoma regions, and then were ranked as order of large liver cell regions and normal liver cell regions. Also the numbers of the positive cells by apoptotic kit tended to be higher in hepatocellular carcinoma regions and cholangiocarcinoma regions but not uniformly in order in all regions and were much less numbers than those of PCNA positive cells. So immunohistochemical methods using PCNA antibody together than using apoptotic kit alone when anti-carcinogen experiments. Rats with positive cells by p53 antibody were 11 of 15 rats(73.4%) in control groups and 12 of 18 rats(66.7%) in CY treated group, respectively. These positive cells appeared focally in early vacuole-occurring regions and were very low in numbers.

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성 hormone이 rat 자궁 발달에 미치는 영향에 대한 proliferating cell nuclear antigen 항체의 면역조직학적 응용 (Immunohistochemical application of anti-proliferating cell nuclear antigen antibody on the development of uterus following sex hormone administration in rats)

  • 고필옥;곽수동
    • 대한수의학회지
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    • 제37권2호
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    • pp.281-289
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    • 1997
  • The study was designed to investigate the effects of progesterone and estrogen on the uterus of rats by immunohistochemical methods using Proliferating Cell Nuclear Antigen (PCNA) antibody. Eighteen female rats(Wistar), weighing initially about 300g, were ovariectomized. These rats were divided into four groups, progesterone-treated group, estrogen-treated group, estrogen+progesterone-treated group, and control group, progesterone-treated group was injected with 1mg of progesterone per rat per day for 2 days and estrogen-treated group with $20{\mu}g$ of $17{\beta}-estradiol$ for 3 days and estrogen+progesterone-treated group with $17{\beta}-estrdiol$ for 3 days and then with progesterone for 2 days as above. In gross findings, the uteri were markedly hypertrophied by estrogen treatment but were not affect in size by progesterone treatment. Immunohistochemical investigation was performed on the cell types with higher appearance of PCNA positive reaction cells in four groups. The groups with higher appearance of the stromal cells were ordered as estrogen-treated group, progesterone-treated group, estrogen+progesterone-treated group, and control group. The muscle cells were ordered as progesterone-treated group, estrogen-treated group, estrogen+progesterone-treated group, and control group. Positive reaction cells of the stromal cells were total 4.6 times higher than those of muscle cells. Therefore, the affect of the hypertrophy on the uterus by estrogen was larger than those of progesterone and affect on the uterus by stromal cells were larger than those of muscle cells. The group with more PCNA positive reaction cells of luminal epithelial cells were ordered as control group, progesterone-treated group, estrogen+progesterone-treated group, and estrogen-treated group, and glandular epithelial cells were ordered as estrogen+progesterone-treated group, progesterone-treated group, control group, and estrogen-treated group. It was suggested that estrogen and progesterone did not affect on the proliferating cells of luminal epithelial cells and affection of progesterone on the development of glandular epithelial cell was larger than that of estrogen.

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Diethylnitrosamine을 투여한 rat 간장의 tumorigenesis에 관하여 2. 종양세포의 apoptosis와 증식에 관한 조직학적 소견 (Diethylnitrosamine-induced hepatic tumorigenesis in rats 2. Histological findings about apoptosis and proliferation of tumor cells)

  • 곽수동;강정부;고필옥
    • 대한수의학회지
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    • 제38권2호
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    • pp.386-393
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    • 1998
  • The study was performed to investigate the histological findings and the appearances of positive cells by immunohistochemical methods using proliferating cell nuclear antigen (PCNA) antibody and apoptotic kit in diethylnitrosamine (DEN) -induced rat liver cancer model. Forty four male rats (Sprague Dawley), initially 5 to 6 weeks of age and 120 to 150gm in body weight were continuously were given with water containing 0.01% DEN for 13 weeks and 3~6 rats per week were randomly sacrified at intervals of a week from 8 weeks to 17 weeks. The interlobular connective tissues in the rat livers were proliferated at early 8 weeks. The vaccuolated or fatty degenerated liver cells were focally distributed and then widely distributed with the passage of weeks and the liver cells with large vacuoles tended to be crowded in focal areas, and the liver cells in some lobules were transformed into small or eosinophilic polyhedral large cells. The hepatocellular carcinoma and the cholangiocarcinoma were simultaneously developed in same liver and tended to be markedly developed after 12 weeks but the development of carcinoma in some livers at same week were less or more advanced as 3~5 week intervals. The regions with more number of positive cells by PCNA antibody or apoptotic kits in livers were ranked as following order ; small hepatocellular carcinoma regions, cholangiocarcinoma regions, trabecular or acinar type carcinoma regions, and large liver cell regions. The numbers of the positive cells by PCNA antibody were more numerous than those by apoptotic kit. So these findings suggested that the volumes and weights of the livers were increasing by more many proliferating of carcinoma cells on the above ordered regions.

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염증성 치은에서 Proliferating Cell Nuclear Antigen(PCNA),${\alpha}$-1-antichymotrypsin, Fibronectin, Transqlutarninase의 분포에 관한 면역조직화학적 연구 (AN IMMUNOHISTOCHEMICAL STUDY ON PROLIFERATING CELL NUCLEAR ANTIGEN, ${\alpha}$-1-ANTICHYMOTRYPSIN, FIBRONECTIN, TRANSGLUTAMINASE IN INFLAMMED GINGIVA)

  • 김재현;유형근;김성호
    • Journal of Periodontal and Implant Science
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    • 제25권2호
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    • pp.253-266
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    • 1995
  • Recently, available interests concerning the biologic significance of the extracellular matrix and proliferating cells associated with periodontal disease has been increased. The distribution or expression of cellular proliferation by PCNA, macrophage detection by ${\alpha}$-l-antichymotrypsin, fibronectin playing a important role in host defence mechanisms indirectly, and transglutaminase that cross linked to fibronectin and stimulate fibrin stabilization were studied in inflammed and healthy gingiva. The excised tissue samples were fixed neutral formalin for 24 hours, embedded with paraffin, sectioned at 4-61lffi in thickness, and immunohistochemically processed by LSAB method. The positive reaction to PCNA was localized in the suprabasal and basal layer of inflammed gingiva and an increasing reactivity was observed than healthy gingiva. ${\alpha}$-I-antichymotrypsin positive cells were localized in the basal layer of inflammed gingiva, and there was no or rare positive cells in healthy gingiva. The positive reaction to fibronectin in inflammed gingiva was more than healthy gingiva,"and shown in the connective tissue subjacent to basement membrane of epithelium and in the periphery of the collagen fiber bundles. The positive cells by transglutaminase in inflammed gingiva were noted in suprabasal, spinous, and keratin layer of epithelium, and slightly increased in the capillaries of connective tissues. But the results of this study demonstrated in vitro reaction. Therefore, the role of PCNA,${\alpha}$-l-antichyrnotrypsin, transglutaminase, fibronectin and coefficient with other growth factor and extracellular matrix were further investigated in vivo.

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Urethane으로 유발된 생쥐 폐샘암종 발생과정에서 Caspase 3, Survivin과 p53 단백 발현 (Expression of Caspase 3, Survivin, and p53 Protein in Urethane Induced Mouse Lung Carcinogenesis)

  • 신종욱;이수환;박언섭
    • Tuberculosis and Respiratory Diseases
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    • 제63권3호
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    • pp.251-260
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    • 2007
  • 연구배경 및 목적: ICR계의 생쥐에 Urethane을 투여하여 발생되는 폐 병변의 형태변화를 관찰하고, 폐 샘암종으로 진행과정에서 세포증식능과 세포자멸사, 세포자멸사와 관련된 조절인자인 caspase3, 자멸사 억제인자인 survivin, 그리고 종양억제 유전자 산물인 p53 단백질이 발암과정에서 발현되는 양상을 관찰함으로써 발암과정에서의 역할을 규명하고자 하였다. 방법: Urethane을 ICR 생쥐에게 복강 내 주사를 하였고, 5주, 15주, 25주에 폐병변을 육안적으로 헤마토실린 및 에오신 염색을 관찰하였고, 면역조직화학적 방법으로 PCNA지수, 세포자멸사 지수, capase 3, survivin 및 p53의 발현을 관찰하였다. 결과: Urethane 투여 5주부터 폐의 증식이 관찰되었고, 샘종은 10주 이후부터 출현하여 시간 경과에 따라 크기와 구조적 변화가 동반되었고, 세포학적 이상소견과 더불어 주변으로 침윤성 변화가 있는 샘암종은 25주 이후에 출현하였다. 세포증식능과 세포자멸사 지수는 폐의 증식증에서는 9.6%와 0.24%, 샘종에서는 23.2%와 1.25%, 그리고 샘암종에서는 55.7%와 5.21%이었다. 따라서 세포증식능과 세포자멸사는 폐 샘암종 발생초기부터 통계적으로 유의하게 지속적으로 증가하였고, 특히 샘암종으로 진행할 경우 현저하게 증가하였다. caspase 3는 증식증에서는 15%, 샘종은 16%의 발현율을 보이는 반면, 폐샘암종은 46.7%의 발현율을 보이면서 암 단계에서 현저하게 발현이 증가하였다. Survivin 단백의 발현은 폐의 증식증, 샘종, 그리고 샘암종으로 진행할수록 발현빈도가 통계적의로 유의하게 증가하였다. p53 단백은 폐의 증식증과 샘종에서는 전혀 발현되지 않았으나 침윤성 샘암종의 일부에서 발현되었다. 이상의 결과로 생쥐의 폐샘암종 발생과정에서 세포증식능과 세포자멸사는 종양의 발생과 진행과정에서 지속적으로 관여함을 알 수 있었다. 또한 survivin 발현은 샘암종의 초기단계에서부터 지속적으로 관여하며, p53 유전자 변이는 초기보다는 암종으로 형질전환이 일어난 후에 부분적으로 발생하는 것으로 보인다. 결론: 본 연구를 통하여 PCNA와 세포자멸사 지수는 암세포로 형질전환하는 위험도를 평가하는 지표로 유용할 것으로 보이며, caspase, survivin과 p53는 urethane에 의해 유도된 생쥐 폐암 모델에서 발암과정에 중요하게 관여하는 단백질로 보인다.

경피적 폐생검의 진단성적 및 합병증 (The Diagnostic Yield and Complications of Percutaneous Needle Aspiration Biopsy for the Intrathoracic Lesions)

  • 장승훈;김철현;고원중;유철규;김영환;한성구;심영수
    • Tuberculosis and Respiratory Diseases
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    • 제43권6호
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    • pp.916-924
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    • 1996
  • 연구배경: 경피적 폐생검은 흉부 병소의 진단을 얻기위해 흔히 이용되는 검사 방법으로써, 축적된 경험, 검칩의 개발, 영상 유도 방법의 개선으로 진단 성적이 높아지고 있으며, 기존의 논문들이 28 - 98%의 진단 성적을 보고하며 시술에 의한 합병증은 그 정도가 가벼워서 비교적 안전하게 시행될 수 있다고 알려져 있다. 저자들은 서울대학병원에서 1년 동안 시행된 경피적 폐생검 결과를 검토하여 그의 진단 성적과 합병증의 빈도와 정도를 조사하고 진단 성적에 영향을 미치는 요인들을 평가하고자 본 연구를 시작하였다. 방법: 1994 년 1월부터 1994년 12월까지, 흉부병소를 가진 236명의 환자에게 시행된 287 회의 경피적 폐생검 결과를 검토하여 양성 및 악성 질환에 대한 진단 성적과 합병증의 발생 빈도를 조사하였다. 병소는 21- 23 G Chiba needle로 흡인하였고, 필요시 19 - 20 G Biopsy gun으로 생검을 실시 하여 미생물학적 검사, 세포진 검사, 병리 조직학적 검사를 시행하였다. 그 결과 및 합병증의 발생 빈도는 병록지만을 참고하여 산출되었다. 병변의 형태와 크기에 따른 진단 성적은 chi square 방법으로 통계 처리하여 비교하였다(P<0.05). 결과: 병변의 위치는 우상엽 26.3%, 우중엽 6.4%, 우하엽 21.2%, 좌상엽 16.8%, 좌하엽 10.6%, 2엽 이상의 부위에 병변이 있었던 경우가 17.4%, 종격동 1.3% 였다. 병변의 형태는 경화성 병변이 19.9%, 결절 또는 종괴성 병변이 80.1% 였다. 236명 중 양성 질환자는 74명, 악성질환자는 142명, 경피적 폐생검을 포함한 어떤 검사로도 진단이 밝혀지지 않은 환자는 22명 이었고, 2명은 폐암과 폐결핵을 같이 가지고 있었다. 236 명을 대상으로 총 287 회의 경피적 폐생검이 시행되었는데, 이것으로 확진된 경우가 양성 질환자의 경우 46명으로 62.2%, 악성 질환자의 경우 117명으로 82.4%의 진단 성적을 보였다. 경피적 폐생검을 1차로 시행하여 진단에 이르지 못한 경우 2차, 3차 재 시술 함으로써 양성 질환의 경우 44.6%, 60.8%, 62.2%로, 악성 질환의 경우 73.9%, 8 1.7%, 82.4%로 진단 성적을 높일 수 있었다. 악성 질환자 43 명과 양성 질환자 9명에서 개흉술을 실시하였는데 개흉술과 경피적 폐생검의 병리학적 검사 결과가 일치하였던 경우는 악성과 양성에서 각각 25 명과 4명 으로 58.1% 와 44.4 %의 일치율을 보였다. 또한 약성 질환자 4명과 양성 질환자 2명은 개흉술 후 악성과 양성의 진단이 바뀌었다. 287례의 시행 중 합병증은 각혈 3 례, 경미한 혈담 55 례, 기흉 36례, 발열 3례로 빈도는 각각 1.0%, 19.2%, 12.5%, 1.0 % 였다. 합병증 발생시 각혈과 혈당은 모두 치료를 요하지 않았고, 기흉은 8례에서 흉관 또는 픽테일 카테터를 삽입하였으며, 발열은 모두 48 시간 내에 자연 소실되었다. 병변의 형태와 크기에 따른 진단 성적의 차이는 없었다. 결론: 경피적 폐생검은 진단 성적이 비교적 높고 합병증의 정도가 낮아 흉부 영소의 진단에 유용하지만 병리 조직학적 진단의 정확도는 개선이 필요하다고 생각된다.

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2-Bromopropane에 의한 유발된 Sprague-Dawley 랫트의 고환위축의 병리학적 관찰 및 Flow Cytometry를 이용한 검사 (Histopathological Observation and Flow Cytometry Analysis of Testicular Atrophy Induced by 2-Bromopropane On the Sprague-Dawley Rat)

  • 손화영;강부현;조성환;차신우;노정구
    • Toxicological Research
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    • 제14권2호
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    • pp.143-149
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    • 1998
  • This study was carried out to evaluate the testicular toxicity of 2-bromopropane (2-BP), which recently caused occupational intoxication on the reproductive and hematopoietic system in Koreans, using light microscopy, immunohistochemistry and flow cytometry. 10 weeks old male Sprague-Dawley (SD) rats were treated with 0.5 g/㎏/day of 2-BP orally for 8 consecutive weeks. The testes of the rats were vascularly perfused with Karnovsky's solution or immersed in Bouin's solution, embedded in plastic and evaluated with light microscopy. And relative proportions of haploid, diploid, and tetra-ploid states of DNA ploidy in the testicular cell suspensions of the SD rats were examined by flow cytometry. 2-BP induced severe testicular atrophy, depletion and degeneration of spermatogonia, spermatocytes, and spermatids and mild hyperplasia of Leydig cells without significant morphological changes. The Leydig cell hyperplasia was confirmed by immunohistochemistry using proliferating cell nuclear antigen (PCNA). The immunopositive cells against PCNA were observed in the nuclei oj some interstitial cells. Relative proportions of haploid states of DNA ploidy decreased in the atrophic testicular cell suspensions comparing with those of the control. In conclusion, 2-BP induced testicular atrophy with Leydig cell hyperplasia as examined by histopathology, immunohistochemistry and DNA flow cytometry.

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