• 대한두경부종양학회지
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• 제20권2호
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• pp.135-142
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• 2004

Objectives: The purpose of this study was to evaluate for prognostic significance of VEGF expression and tumor angiogenesis in papillary carcinomas of the thyroid. Materials and Methods: The materials were 79 cases of papillary thyroid carcinomas, and age, sex, tumor size, multiplicity of tumor, capsular invasion, lymph node metastasis, recurrence, TNM stage, DeGroot stage and AMES scale were evaluated. An immunohistochemical stains for CD 34 to estimate microvessel density (MVD), and VEGF were done. MVD was defined as an average count of vessels per ${\times}400$ power field in the most vascularized area. VEGF expression was interpreted as 1+ and 2+ according to staining intensity and percentages of positive cells. Results: Mean score of MVD was $39.7{\pm}16.9.$ MVD were significantly higher in cases with capsular invasion (p=0.0001), lymph node metastasis (p=0.0001), TNM stage III (p=0.0022), DeGroot stage III (p=0.0163) and high risk group by AMES scale (p=0.0001). VEGF 2+ expression rate was significantly increased in cases with capsular invasion and lymph node metastasis (p=0.0006, p=0.0013), and in cases with TNM stage III, DeGroot stage III and high risk group by AMES scale (p=0.0236, p=0.0003, p=0.0293). In VEGF 2 + expression group, MVD was significantly higher than in VEGF 1 + group (p=0.0008), and MVD showed positive relation to VEGF 2 + expression (r=0.4616). Conclusion: VEGF expression and high MVD were significantly correlated to capsular invasion, lymph node metastasis, TNM stage III, DeGroot stage III and high risk group by AMES scale. The expression of VEGF and high MVD could be considered to be one of prognostic factor in papillary thyroid carcinomas.

• Applied Microscopy
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• 제17권1호
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• pp.47-64
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• 1987

Morphological difference of the renal glomerulus at different age groups have been studied in young (three month-old), adult (twelve month-old) and old (thirty month-old) Fisher strain 344 rats. Pieces of the tissues were taken from renal corticies prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1 M Millonig's phosphate buffer, pH 7.3), following by post-fixation with 1% osmium tetroxide (0.1 M Millonig's phosphate buffer, pH 7.3) and embedded within Araldite. The ultrathin sections contrasted with uranyl acetate and lead citrate were observed under a JEM 100CX electron microscope. The mean thickness of glomerular basal lamina and Bowman's capsule were determined by measuring the thinnest portion of basal lamina, and by taking the average of 50 readings from electron micrographs at different ages. The numerical changes of the slit pores were compared based upon the numbers over the length of 10um of glomerular basal lamina. The results were as follow: 1. The thickness of glomerular basal lamina is increased during aging; 140.4 nm in young rats, 270.0 nm in adult ones, and 437.8 nm in old ones. 2. The thickness of basal lamina of parietal cells of Bowman's capsule is 187.5 nm in young rats, 914.0 nm in adult ones, and 2850.0 nm in old ones. 3. The numbers of the slit pores of basal lamina are reduced during aging, 30.3 slit pores/$10{\mu}m$ in adult ones, and 24.2 slit pores/$10{\mu}m$ in old ones. 4. Accumulation of dense intracytoplasmic filamentous material in the parietal cells of Bowman's capsule is increased in the vicinity of the basal lamina during aging. The proximal tubule-like epithelial cell in Bowman's capsule is observed at one glomerulus in a young rat. 5. The endothelial cells are edematous and form balloon-like structure protruding into capillary lumen in young and old rats. 6. Cytoplasm of the podocyte shows a variety of alteration during aging, such as swelling of mitochondria and of endoplasmic reticulum, and increase of microtubules, microfilaments, lysosomes and lamellated myelin structures, etc. Accumulation of dense intracytoplasmic material in the foot processes is increased in the vicinity of the basal lamina during aging. The podocytic membrane-like structures are seen in young and o]d rats. 7. The mesangial matrices and mesangial cells are increased during aging, and slight swelling of endoplasmic reticulum and Golgi cisternae in young and old rats.

• Parasites, Hosts and Diseases
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• 제49권2호
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• pp.115-123
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• 2011

Toxoplasma gondii Korean isolate (KI-1) tachyzoites were inoculated intraduodenally to BALB/c mice using a silicon tube, and the course of infection and immune responses of mice were studied. Whereas control mice, that were infected intraperitoneally, died within day 7 post-infection (PI), the intraduodenally infected mice survived until day 9 PI (infection with $1{\times}10^5$ tachyzoites) or day 11 PI (with $1{\times}10^6$ tachyzoites). Based on histopathologic (Giemsa stain) and PCR (B1 gene) studies, it was suggested that tachyzoites, after entering the small intestine, invaded into endothelial cells, divided there, and propagated to other organs. PCR appeared to be more sensitive than histopathology to detect infected organs and tissues. The organisms spread over multiple organs by day 6 PI. However, proliferative responses of splenocytes and mesenteric lymph node (MLN) cells in response to con A or Toxoplasma lysate antigen decreased significantly, suggesting immunosuppression. Splenic $CD4^+$ and $CD8^+$ T-Iymphocytes showed decreases in number until day 9 PI, whereas IFN-${\gamma}$ and IL-10 decreased slightly at day 6 PI and returned to normal levels by day 9 PI. No TNF-${\alpha}$ was detected throughout the experimental period. The results showed that intraduodenal infection with KI-1 tachyzoites was successful but did not elicit significant mucosal immunity in mice and allowed dissemination of T. gondii organisms to systemic organs. The immunosuppression of mice included reduced lymphoproliferative responses to splenocytes and MLN cells to mitogen and low production of cytokines, such as IFN-${\gamma}$, TNF-${\alpha}$, and IL-10, in response to T. gondii infection.

• Radiation Oncology Journal
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• 제34권3호
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• pp.223-229
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• 2016

Purpose: This study is to investigate the effect of captopril when combined with irradiation. Materials and Methods: 4T1 (mouse mammary carcinoma) cells were injected in the right hind leg of Balb/c mice. Mice were randomized to four groups; control (group 1), captopril-treated (group 2), irradiated (group 3), irradiated and captopril-treated concurrently (group 4). Captopril was administered by intraperitoneal injection (10 mg/kg) daily and irradiation was delivered on the tumor-bearing leg for 15 Gy in 3 fractions. Surface markers of splenic neutrophils (G-MDSCs) and intratumoral neutrophils (tumor-associated neutrophils [TANs]) were assessed using flow cytometry and expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor 1 alpha ($HIF-1{\alpha}$) of tumor was evaluated by immunohistochemical (IHC) staining. Results: The mean tumor volumes (${\pm}$standard error) at the 15th day after randomization were $1,382.0({\pm}201.2)mm^3$ (group 1), $559.9({\pm}67.8)mm^3$ (group 3), and $370.5({\pm}48.1)mm^3$ (group 4), respectively. For G-MDSCs, irradiation reversed decreased expression of CD101 from tumor-bearing mice, and additional increase of CD101 expression was induced by captopril administration. Similar tendency was observed in TANs. The expression of tumor-necrosis factor-associated molecules, CD120 and CD137, are increased by irradiation in both G-MDSCs and TANs. Further increment was observed by captopril except CD120 in TANs. For IHC staining, VEGF and $HIF-1{\alpha}$ positivity in tumor cells were decreased when treated with captopril. Conclusion: Captopril is suggested to have additional effect when combined to irradiation in a murine tumor model by modulation of MDSCs and angiogenesis.

• Journal of Chest Surgery
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• 제33권6호
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• pp.494-501
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• 2000

Background: Transmyocardial laser revascularization(TMLR) for revascularizing ischemic myocardium in patients was originally based on the assumption that laser channels remain their patency much longer. But recent studies show that laser channels did not remain open and that TMLR could achieve treatment benefits without long-term channel patency. The angiongencesis is currently thought to be induced by non-specific inflammatory response to mechanical tissue injury. This study is to evaluate hypothesis that various transmyocaridal mechanical revascularization(TMMR) may induce the angiogenic responses similar to that seen with TMLR, and transmyocaridal polymer stent revascularization(TMSR), the polymer stent in the myocardial tissue is hydrolyzed in 2 weeks, may enhance the non-specific inflammatory reaction resulting angiogenesis. Furthermore, polymer myocaridal stent channels remain long-term patency. Material and Method: Eight domestic pigs underwent ligation of the proximal circumflex artery, and 2 weeks later they were randomized to undergo transmycardial acupunctural revascularization (TMPR, Group I) of the left lateral wall with 18-G needle(n=2), to undergo transmyocardial (TMDR, Group II) with industrial 2mm steel drill(n=2), to undergo transmyocardial polymer stent revascularization (TMSR, Group III) after drilling the infarcted myocardium(n=2), the stent is poly(lactic acid-co-glycolic acid), which is self-degradated in the myocardium, and to a control group the ischemic zone was unterated(n=2). All the pigs were sacrificed after 4 weeks TMMR. Sections from the ischemic zone were submitted for vascular endothelial growth factor (VEGF) ELISA and histology. Result: There were makedly increase in the VEGF immunoassay in the ischemic zone of the TMMR group compared to the ischemic zone of the control group(control: each 30.85 and 43.15pg/mg protein, TMPR: each 44.14 and 68.61 pg/mg protein, TMDR: each 65.92 and 78.65 pg/mg protein, TMSR: each 177.39 and 168.87 pg/mg protein). TMSR channels caused greatest VEGF expression than channels made by other group and the polymer stent channels remained vacuole after 4 weeks. Conclusion: Transmyocardial polymer stent revascularization promoted the most angiogenci response by the VEGF immunoassay, although our study did not show the statistical significancy. The channels remained but the flow patency was not verified. Transmyocardial polymer stent revascularization (TMSR) is desirable in future experimental trials and in view of the significant cost implications comparable to that of laser.

• Journal of Chest Surgery
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• 제37권2호
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• pp.113-118
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• 2004

혈관재건술에 사용되는 냉동 보존된 동종동맥편에 대한 지금까지의 연구는 주로 냉동방법, 냉동전처치, 보존온도에 관심이 집중되어 있었으나, 최근에 해동방법이 균열발생의 중요한 인자가 된다는 연구가 있었다. 이에 저자들은, 같은 조건으로 냉동 보존된 대동맥조직을 서로 다른 두 가지 방법으로 해동시켜, 조직손상 정도를 비교함으로써 이상적인 조직의 해동방법을 알아보고자 하였다. 대상 및 방법: 2,500 g의 토끼에서 대동맥편을 획득한 후, 일반적인 방법에 따라 냉동 보존한 뒤 37$^{\circ}C$의 온수에 급속 해동한 군(RT)과 1$^{\circ}C$/min의 속도로 완속 해동시킨 군(ST)으로 나누고, 각 군에서 apoptosis 발생을 평가하기 위해 각 군의 전체 세포 중 TUNEL (＋) 세포의 비를 비교하였고, 광학현미경하에서 해동된 조직편의 조직학적 특성을 비교 관찰하였다. 결과: 1. TUNEL test상, RT군에서 ST군에 비해 TUNEL (＋) 세포의 비가 유의하게 높았다. 2. 광학현미경하 조직소견상, RT군에서 세포부종과 혈관내피층의 박탈, 소수포 형성, 그리고 이형성세포 등의 소견이 ST군에 비해 더 많이 관찰되었다. 결론: 냉동 보존된 토끼의 대동맥조직에 대한 해동방법의 차이에서, 완속 해동방법이 조직의 형태학적 손상과 apoptosis발생을 감소시켰다. 따라서, 향후 냉동조직을 이용한 이식술에서 술 후 합병증의 발생을 감소시키고 예후를 개선하기 위해서는 기존의 급속 해동하는 방법보다 완속 해동하는 방법의 사용을 고려해야 할 것이다.

• Journal of Chest Surgery
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• 제40권4호
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• pp.256-263
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• 2007

배경: 혈관내막과다증식은 혈관평활근세포가 내막에서 과다증식하여 나타나며, epigallocatechin-3-gallate(EGCG)는 혈관평활근세포의 증식을 억제하는 효과가 있는 것으로 알려져 있다. 따라서 EGCG는 혈관내막과다증식을 억제할 수 있을 것으로 생각된다. 대상 및 방법: 다른 농도의 EGCG를 포함한 배양 배지에서 인간제대정 맥내피세포(HUVEC)와 쥐대동맥평활근세포(RASMC)를 배양하고, 세포증식과 이동속도를 측정하였다. 20마리의 개의 양측 경동맥에 자가경정맥을 이식하였으며, 실험군(n=10)에 대해서는 정맥도관을 이식 전 30분간 EGCG용액에 보관하였으며, 이식 후 300mM의 EGCG를 혈관주위에 도포하였다. 6주 후에 경정맥이식편의 내막과 중간막의 두께를 측정하였다. 결과: 내피세포와 혈관평활근세포의 증식은 EGCG에 의해 억제되었다. 혈관평활근세포의 이동성은 EGCG에 의해 억제되었으나, 내피세포의 이동성은 영향을 받지 않았다. 동물실험 결과 대조군에 비해 EGCG군에서 내막의 두께가 얇았으며(p<0.05), 중간막의 두께는 두 군간에 차이가 얼었고, 내막/중간막의 두께비는 EGCG군에서 낮게 관찰되었다(p<0.05). 결론: EGCG는 혈관수술 후 혈관내막과다증식을 억제하는 효과가 있으며, 이는 혈관평활근세포의 증식 및 이동을 억제하여 효과를 나타낸다고 생각된다. 따라서 EGCG는 혈관내막과다증식을 예방하는 치료에 효과적으로 사용될 수 있을 것으로 생각된다.

• Journal of Periodontal and Implant Science
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• 제27권4호
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• pp.883-908
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• 1997

Bone remodeling is characterized by the coupling of osteoclast-mediated bone resorption and osteoblast-mediated bone formation. The process is tightly regualted at the local level by an incompletely known netwotk of peptide and non-peptide fators. Nitric oxide(NO), synthesized by nitric oxide synthetase(NOS) from L-arginine, is becoming recognized as an important bioregualtory molecule in a variety of tissue, but little is known about its possible role in periodontal tissue. The purpose of this study is to investigate the expression of nitric oxide synthetase(NOS) in inflamed gingiva and the effects of cytokine on the expression of NOS protein. The expression of NOS in gingival tissue was evaluated by immunohistochemical staining for $NOS_1$, $NOS_2$, $NOS_3$. The effect of cytokine on the expression of NOS in human periodontal ligament cells and osteoblast-like HOS cells by western blot analysis. Further, we studied that NO functions in periodontal ligament cells as a regulatory molecule. PDL cells incubated with NOS inhibitor and donor. The protein expression, type I collagen & non-collagenous protein, nitrate production and cell proliferation were evaluated The results were as follows. 1. $NOS_1$, $NOS_2$, $NOS_3$ was rarely distributed in healthy gingiva, but stronger stained in gingival epithelium, endothelial cells, and mononuclear cells of inflammed gingiva. 2. The cytokine stimulated $NOS_1$, and $NOS_3$ protein were not inducing or inhibitory effect to compared with control in PDL and HOS cells. 3.Incubation of cells with combination of $TNF-{\alpha}$, $IFN-{\gamma}$, LPS result in a time dependant increase in $NOS_2$ expression, reaching a maximal level after 24 hours of stimulation. 4. The osteonectin protein inhibitory effect of NMA, inhibitor of NOS, was reversed by Larginine in dose dependant manner. 5. NMA decreased cell poliferation and nitrate production, but the inhibitory efffect of NMA was also prevented by the NO donor, sodium nitropruiside. These results suggest that exogenously synthesized NO was playing a stimulating effect on cell proliferation or on non-collagenous protein expression. Therefore NO have an important role in mediation of localized bone destruction associated inflammatory bone disease such as periodontitis.

• 한국식품영양과학회지
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• 제43권3호
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• pp.374-380
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• 2014

본 연구에서는 야관문 추출물의 마우스 대식세포에 대한 항염증 활성과 창상유발 동물실험 모델을 통한 창상치유 효과를 조사하였다. RAW264.7 세포에서 야관문 추출물은 0.2 mg/mL 이하 농도에서 세포생존에 영향을 주지 않았으며, 염증반응이 활성화된 대식세포에 대해 농도 의존적으로 유의적인 NO 생성 감소를 나타내었다. 창상유발 동물실험 모델에서 야관문 추출물을 함유한 화장품 조성물의 창상치유효과에 대해 육안적으로 관찰한 결과, SCO군과 CCO군보다 야관문 추출물을 함유한 SSP군에서 약 20~30% 빠른 상처면적 감소 효과를 나타내었으며, 반흔 크기 역시 약 12% 작게 형성되었다. 또한 SSP군 조직의 외피와 진피 재생회복속도가 빨라진 것을 Masson's trichrome 염색을 통해 확인할 수 있었으며, VEGF 및 TGF-${\beta}1$ 유전자 발현이 SCO군과 비교 시 각각 감소 및 증가하였다. 이러한 결과는 야관문 추출물이 항염증 및 교원질 생성 유도를 통한 조직재생 활성에 기여하여 창상치유 속도를 가속화하고 반흔 면적을 감소시킬 수 있는 피부 창상치유와 관련한 코스메슈티컬 소재로써 산업적 활용이 가능함을 보여준다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권5호
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• pp.2353-2358
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• 2014

Purpose: We aimed to detect the expression of HIF-1${\alpha}$, VEGF, HPSE-1 and CD31 in SKOV3 xenografts in nude mice treated with different doses of ionizing radiation, trying to explore the possible mechanism of hypoxia and radioresistance. Methods: Nude mice bearing SKOV3 xenografts were randomly divided into 4 groups: Group A (control group, no ionizing radiation), Group B (treated with low dose of ionizing radiation: 50cGy), Group C (treated with high dose of ionizing radiation: 300cGy), Group D ( combined ionizing radiation, treated with ionizing radiation from low dose to high dose : 50cGy first and 300cGy after 6h interval). The mRNA levels of HIF-1 and VEGF in each group were detected by real time polymerase chain reaction, while HPSE-1 expression was measured by ELISA. The microvessel density (MVD) and hypoxic cells were determined through immunohistochemical (IHC) staining of CD31 and HIF-1a. Results: Significant differences of HIF-1${\alpha}$ mRNA level could be found among the 4 groups (F=74.164, P<0.001): Group C>Group A>Group D> Group B. The mRNA level of VEGF in Group C was significantly higher than in the other three groups (t=-5.267, P=0.000), while no significant difference was observed among Group A, B and D (t=1.528, 1.588; P=0.205, 0.222). In addition, the MVD was shown to be the highest in Group C (t=6.253, P=0.000), whereas the HPSE-1 level in Group A was lower than in Group B (t=14.066, P=0.000) and higher than in Group C (t=-21.919, P=0.000), and similar with Group D (t=-2.066, P=0.058). Through IHC staining of HIF-1a, the expression of hypoxic cells in Group A was (++), Group B was (+), Group C was (+++) and Group D was (+). Conclusion: Ionizing radiation with lowerdoses might improve tumor hypoxia through inhibiting the expression of HIF-1 and HPSE-1, whereas higherdoses worsen tumor hypoxic conditions by up-regulating HIF-1${\alpha}$, HPSE-1, VEGF and CD31 levels. A protocol of low-dose ionizing radiation followed by a high-dose irradiation might at least partly improve tumor hypoxia and enhance radiosensitivity.