• 한국가축번식학회지
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• 제21권3호
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• pp.265-274
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• 1997

The purpose of this study was attempted to investigate the a, pp.arences of healthy or artretic follicles in ovaries following repeats of pregnant mare serum gonadotropin(PMSG) treatments for superovulation in nulliparous rats. Thirty two rats(Sprague-Dawely, about 200-250 gm) were randomized into 4 groups. Control group rats were sacrified at estrus phase confirmed by vaginal smear. PMSG-treated group 1 rats, PMSG-treated group 2 rats and PMSG-treated group 3 rats were sacrified at 48 hrs after injection once with PMSG 25 IU, after 2 repeated injection by a week interval, and 3 repeated injection, respectively. The ovaires of rats were removed and then sections by paraffin embedding were stained with H-E or immunohistochemical staining using proliferating cell nuclear antigen monoclonal antibody (PCNA m Ab) and apoptotic kit. The criteria of follicle classification was based as small follicles with preantral follicles with 2~4 layers of granulosa cells surrounding the oocyte, as secondary follicles with more than 5 layers of granulosa cells and early signs of antral cavity or with small clefts on either side of the oocytes, and as tirtiary follicles with a single medium sized antral cavity or large well-formed antral cavity, respectively. The proportions of atretic follicles from small and middle follicles in immunohistochemical staining using PCNA m Ab were 17.9% and 21.3% in control group, 15.5% and 23.5% in PMSG-treated group 1, 24.3% and 26.7% in PMSG-treated group 2, 18.1% and 30.2% in PMSG-treated group 3, respectively. Groups with atretic follicles of higher proportion were ordered as PMSG-treated group 3, PMSG-treated group 2, PMSG-treated group 1 and control group. The proportions of positive cells in small, middle and large follicles were 31.1%, 33.5% and 28.5% respectively. The follicles with positive cells of higher proportion were ordered middle, small and large follicles. In immunohischemical staining using apoptotic kits, small follicles in all 4 groups did not contain positive cells, and proportions of atretic follicles from middle and large follicles were 24.9, 30.7, 33.8 and 40.1% in control, PMSG-treated gruop 1, PMSG-treated group 2 and PMSG-treated group 3, respectively. These results suggested that repeats of PMSG treatment increased proportion of atretic follicles in ovaries, and middle follicles are more quickly developing than small or large follicles.

• 대한약침학회지
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• 제17권1호
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• pp.27-34
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• 2014

Objectives: This experiment was conducted to examine the toxicity of Water soluble Carthmi-Flos herbal acupuncture (WCF) by administering a single intramuscular dose of WCF in 6-week-old, male and female Sprague-Dawley rats and to find the lethality dose for WCF. Methods: The experiment was conducted at Biotoxtech according to Good Laboratory Practices under a request by the Korean Pharmacopuncture Institute. This experiment was performed based on the testing standards of "Toxicity Test Standards for Drugs" by the Ministry of Food and Drug Safety. Subjects were divided into 4 groups: 1 control group in which normal saline was administered and 3 test groups in which 0.1, 0.5 or 1.0 mL of WCF was administered; a single intramuscular dose was injected into 5 males and 5 females in each group. General symptoms and body weights were observed/measured for 14 days after injection. At the end of the observation period, hematological and clinical chemistry tests were performed, followed by necropsy and histopathological examinations of the injected sections. Results: No mortalities were observed in any group. Also, symptoms, body weight, hematology, clinical chemistry and necropsy were not affected. However, histopathological examination of the injected part in one female in the 1.0-mL group showed infiltration of mononuclear cells and a multi-nucleated giant cell around eosinophilic material. Conclusion: Administration of single intramuscular doses of WCF in 3 groups of rats showed that the approximate lethal dose of WCF for all rats was in excess of 1.0 mL, as no mortalities were observed for injections up to and including 1.0 mL.

• 한국안광학회지
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• 제18권4호
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• pp.533-540
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• 2013

목적: 본 연구에서는 발생 중[태어난 지 5일(P5), 10일(P10)] 마우스 망막의 ON-OFF 방향특이성 신경절세포 수상돌기 상에서 NMDA R1 수용체의 시냅스 패턴을 연구하고자 하였다. 방법: ON-OFF 방향특이성 신경절세포는 Lucifer yellow를 주사하여 형태학적 특징으로 동정하였다. 이극세포로부터의 글루타메이트의 흥분성 유입을 확인하기 위해 membrane traffic motor 단백질 마커인 kinesin을 이용하였다. 결과: 본 연구에서 P5, P10의 ON-OFF 방향특이성 신경절세포를 동정할 수 있었으며, NMDA R1의 면역반응반점은 내망상층에서 강하게 나타났다. ON-OFF 방향특이성 신경절세포의 수상돌기상의 수용체의 분포패턴에서 방향특이성을 예측할 수 있는 어떠한 비대칭성도 발견하지 못하였다. 결론: 이극세포로부터의 글루타메이트성 자극유입은 P5, P10 단계에서 모두 균형적으로 유입되며, 방향특이성은 NMDA R1 수용체의 특이적인 패턴에 있지 않음을 밝혔다.

• The Korean Journal of Physiology and Pharmacology
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• 제26권6호
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• pp.427-438
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• 2022

Pyroptosis, a form of cell death associated with inflammation, is known to be involved in diabetic nephropathy (DN), and discoid domain receptor 1 (DDR1), an inflammatory regulatory protein, is reported to be associated with diabetes. However, the mechanism underlying DDR1 regulation and pyroptosis in DN remains unknown. We aimed to investigate the effect of DDR1 on renal tubular epithelial cell pyroptosis and the mechanism underlying DN. In this study, we used high glucose (HG)-treated HK-2 cells and rats with a single intraperitoneal injection of streptozotocin as DN models. Subsequently, the expression of pyroptosis-related proteins (cleaved caspase-1, GSDMD-N, Interleukin-1β [IL-1β], and interleukin-18 [IL-18]), DDR1, phosphorylated NF-κB (p-NF-κB), and NLR family pyrin domain-containing 3 (NLRP3) inflammasomes were determined through Western blotting. IL-1β and IL-18 levels were determined using ELISA. The rate of pyroptosis was assessed by propidium iodide (PI) staining. The results revealed upregulated expression of pyroptosisrelated proteins and increased concentration of IL-1β and IL-18, accompanied by DDR1, p-NF-κB, and NLRP3 upregulation in DN rat kidney tissues and HG-treated HK-2 cells. Moreover, DDR1 knockdown in the background of HG treatment resulted in inhibited expression of pyroptosis-related proteins and attenuation of IL-1β and IL-18 production and PI-positive cell frequency via the NF-κB/NLRP3 pathway in HK-2 cells. However, NLRP3 overexpression reversed the effect of DDR1 knockdown on pyroptosis. In conclusion, we demonstrated that DDR1 may be associated with pyroptosis, and DDR1 knockdown inhibited HG-induced renal tubular epithelial cell pyroptosis. The NF-κB/NLRP3 pathway is probably involved in the underlying mechanism of these findings.

• 대한골관절종양학회지
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• 제20권1호
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• pp.14-21
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• 2014

목적: 본 교실에서 경험한 랑게르한스 세포 조직구증의 임상적 특징을 알아보고 치료방법 및 예후를 알아보고자 한다. 대상 및 방법: 1996년 8월부터 2013년 6월까지 본 교실에서 수술 후 랑게르한스 세포 조직구증으로 확진 된 총 28례를 대상으로 발병 당시부터 최근까지의 진찰소견, 의무기록, 방사선 사진, 병리소견을 토대로 임상소견과 진단 및 치료결과에 대해 후향적으로 분석하였다. 결과: 총 28례로 소아 22례 성인 6례였다. 발병나이는 0.6세에서 51세 사이에서 발생하였으며 평균연령은 14.8세였다. 추시 기간은 6개월에서 134개월까지 평균 44.6개월 이었다. 환자의 구성은 남자 20명 여자 8명으로 남녀성비는 2.5:1이었다. 초기증상은 동통 18례, 병적 골절 5례, 종괴 3례, 방사선 검사상 우연히 발견된 경우 1례, 사경 1례였다. 방사선학적 소견에서는 모든 증례에서 골 용해 소견을 보였으며 이중 13례에서는 골막 반응이 동반되었으며 1례에서는 연부조직 침범이 관찰되었다. 임상적 분류로 28례 모두 호산구성 육아종 이었으며 조직구 협회가 제시한 분류법으로는 단일계통의 다발 병소 질환 3례, 단일 병소 질환 25례였다. 전례에서 조직 생검술이 시행되었으며 6례에서는 생검술 이후 특별한 치료 없이 골 유합을 얻었으며 11례에서는 스테로이드 국소 주입술을 시행하였으며 11례는 조직생검과 동시에 소파술 및 골이식술을 시행하거나 추가로 금속 내고정술을 시행하였다. 추시기간 중 환자가 사망한 경우는 없었다. 병변이 국소 재발한 경우는 없었으나 신체 다른 부위에 골병변이 생긴 경우가 3례 있었다. 합병증은 총 2례가 있었으며 소파술 및 골 이식술을 시행한 이후 감염소견을 보여 변연절제술을 시행한 경우였다. 결론: 랑게르한스 세포 조직구증 환자에서 급격한 전신적 발병을 가져오는 경우는 매우 드물기 때문에 초기 진단 시에 어린 환아에게 과도한 방사선 노출을 가져오는 검사는 지양해야 할 것으로 생각된다. 질환의 확진을 위해서는 조직 생검이 필요하며 조직학적 확진 이후에는 경과 관찰만으로도 대부분에서 만족스러운 결과를 얻을 수 있으나 술 전 병적 골절이 있거나 골 파괴 병변의 범위가 넓은 경우 소파술 및 골 이식술과 내고정술이 필요할 수도 있다. 3-6개월간의 경과관찰에도 병변의 호전이 느리거나 일상생활에 지장을 주는 통증을 호소하는 경우 스테로이드 국소 주입법이 좋은 치료가 될 수 있다.

• Tuberculosis and Respiratory Diseases
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• 제40권2호
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• pp.104-111
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• 1993

연구배경 : 그람 음성균의 내독소는 ARDS를 유발하는 제일 흔한 원인으로 알려져 있으며 그 기전중에 내독소에 의해서 폐로의 유입이 증가되고 활성화된 호중구에서 분비하는 산소기가 증가하여 모세혈관 내피세포등에 손상을 입히기 때문이라는 설이 유력하다. 그러한 증가된 산소기를 처리하는 항산화효소가 내독소를 대량 투여하거나 소량 반복투여시 어떻게 변화하는지 알아보고자 본 실험을 하였다. 방법 : 250~300g 되는 쥐에 내독소 대량투여군은 7mg/kg body weight로 내독소를 1회 복강내 주사하였고 소량 반복투여군은 1 mg/kg body weight로 10일간 매일 복강내 주사하였다. 대량투여군은 주사후 6, 12, 24시간후에, 소량 반복투여군은 주사후 3, 7, 10일에 각각 기관지 폐포세척술을 시행하고 살해하였다. 폐동맥을 통하여 생리 식염수를 주입하여 적혈구를 제거한후 양측 폐를 들어내고 무게를 잰후 무게의 5배되는 용량의 50 mM potassium phosphate 완충액하에 3분 간 homogenize시켰다. 이들 $4^{\circ}C$ 에서 60분간 100,000g의 속도로 원심분리하여 상충액을 $-70^{\circ}C$에 보관하였다가 Cu, Zn SOD, Mn SOD, catalase, GSH-Px와 albumin 농도를 측정하였다. 결과: 1) 폐의 wet/dry 무게비 및 기관지 폐포세척액내 albumin 농도는 내독소 대량투여후 12시간에 최고로 증가하였고 기관지 폐포세척액내 호중구는 내독소 대량투여후 6 시간에 최고로 증가하였다. 2) Cu, Zn SOD (IU/mg protein)는 대조군 ($66.7{\pm}26.3$)에 비해 대량투여군에서 6시간후 ($21.86{\pm}5.79$)와 12시간후 ($31.96{\pm}9.54$)에 각각 의의있는 감소를 나타냈으며 소량 반복투여군에서는 큰 변화가 없었다. 3) Mn SOD, catalase, GSH-Px의 양은 대량 또는 소량투여군 모두에서 특별한 변화를 관찰할 수 없었다. 결론: 내독소를 대량투여시 급성 폐부종의 소견과 함께 폐조직내 Cu, Zn SOD가 6시간 및 12시간후에 현저히 감소함을 관찰할 수 있었는데 이러한 현상은 내독소에 의해서 폐의 세포가 손상을 받은 때문으로 생각된다. 따라서 SOD를 외부에서 보충해주거나 폐내에서 합성을 증가시키는 방법으로 급성 폐손상의 진행을 막을수 있는지에 대한 연구가 진행되어야 할 것으로 사료된다.

• 대한핵의학회지
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• 제33권2호
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• pp.120-130
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• 1999

목적: 갑상선기능항진증 환자에서 병소의 국소화를 위하여 시행하는 Tc-99m MIBI를 이용한 double-phase 부갑상선 신티그라피와 SPECT 방법의 진단적 유용성을 평가하고, 부갑상선의 호산성 세포량과 Tc-99m MIBI 섭취와의 관계를 알고자 하였다. 대상 및 방법: 임상적으로 부갑상선기능항진증이 의심되어 부갑상선 신티그라피가 시행된 16명의 환자중 10명에서 수술이 시행되었고, 여기서 얻은 28개의 부갑상선 조직들을 대상으로 분석하였다. Tc-99m MIBI를 환자에게 정맥주사 후 15분과 2시간에 조기와 지연 바늘구멍 영상을 얻었고 이어서 SPECT 영상을 촬영하였다. 수술로 제거된 부갑상선 조직의 무게와 호산성 세포량을 구하였고, 신티그라피소견을 병리조직 결과와 비교하였다. 결과: 수술로 6개의 부갑상선종과 9개의 부갑상선 비대증이 진단되었다. 조기와 지연 바늘구멍 영상의 민감도, 특이도, 그리고 양성 예측률은 각각 46.7% (7/15),76.9% (10/13), 70% (7/10)과 66.7% (10/15), 92.3% (12/13), 90.9% (10/11)이었다. SPECT 영상은 바늘구멍 영상에 비해 1개의 부갑상선 증식증 병소를 더 진단할 수 있었다. 전체적으로 조기와 지연 바늘구멍 영상과 SPECT 영상을 종합하면 민감도, 특이도, 그리고 양성 예측률은 73.3% (l1/15), 100% (13/13), 100% (11/11)였다. 부갑상선기능항진증의 원인 중부갑상선종은 민감도가 100% (6/6)였으나 부갑상선 증식증은 55.5% (5/9)이었다. 호산성 세포량은 부갑상선종과 부갑상선 증식증에서 정상 부갑상선에 비해 증가되었으나(p<0.0001) 부갑상선종과 부갑상선증식증을 비교하면 호산성 세포량과 무게에는 유의한 차이는 없었다. 결론: Tc-99m MIBI를 이용한 double-phase 부갑상선 신티그라피와 SPECT는 부갑상선기능항진증 환자의 병소 국소화에 효과적인 방법으로 특히 부갑상선종의 국소화에 유용하였다. 부갑상선종과 부갑상선 증식증에서 모두 호산성 세포량이 증가되어 있었으나, 부갑상선종에서는 민감도가 높았고, 부갑상선 증식증에서는 낮았다. 부갑상선증식증에서 민감도가 낮은 원인에 대해서는 더 연구가 필요할 것으로 생각된다.

• BMB Reports
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• 제28권3호
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• pp.221-226
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• 1995

Dopamine mediates inhibitory responses in Helix aspersa neurons from the right parietal lobe ("F-lobe") of the circumoesophageal ganglia. The effects appeared as a dose-dependent hyperpolarization of the plasma membrane and a decrease in the occurrence of spontaneous action potentials. The average hyperpolarization with 5 ${\mu}m$ dopamine was -12 mV (${\pm}1.5$mV, S.D., n=12). Dopamine also modulated the currents 'responsible for shaping the action potentials in these neurons. When dopamine was added and action potentials were triggered by an injection of current, the initial depolarization was slowed, the amplitude and the duration of action potentials were decreased, and the after-hyperpolarization was more pronounced. The amplitude and the duration of action potential were reduced about 15 mV and about 13% by 5 ${\mu}m$ dopamine, respectively. The effects of dopamine on the resting membrane potentials and the action potentials of Helix neurons were dose-dependent in the concentration range 0.1 ${\mu}m$ to 50 ${\mu}m$. In order to show 1) that the effects of dopamine were mediated by dopamine receptors rather than by direct action on ionic channels and 2) which type of dopamine receptor might be responsible for the various effects, we assayed the ability of mammalian dopamine receptor antagonists, SCH-23390 (antagonist of D1 receptor) and spiperone (antagonist of D2 receptor), to block the dopamine-dependent changes. The D1 and D2 antagonists partially inhibited the dopamine-dependent hyperpolarization and the decrease in action potential amplitude. They both completely blocked the decrease in action potential duration and the increase in action potential after-hyperpolarization. The dopamine-induced slowdown of the depolarization in the initial phase of the action potentials was less effected by SCH-23390 and spiperone. From the results we suggest 1) that Helix F-lobe neurons may have a single type of dopamine receptor that binds both SCH-23390 and spiperone and 2) that the dopamine receptor of Helix F-lobe neurons may be homologous with and primitive to the family of mammalian dopamine receptors.

• 대한수의학회지
• /
• 제26권1호
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• pp.117-124
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• 1986

The present study was undertaken to evaluate rosette formation and NK activity of splenic lymphocytes in 3-methylcholanthrene (MCA) treated mice. Mice were sensitized iv with 0.1ml of 1% sheep red blood cell (SRBC) suspension were treated with a single ip injection of olive oil alone or with different doses of MCA in oil at various time before or after sensitization, and were challenged at 4 days after SRBC. Rosette formation and NK activity of splenic lymphocytes were measured at 24 hours after challenge. Erythrocyte(E) rosette formation of splenic lymphocytes was significantly depressed in mouse treated with large dose of MCA (5~50mg) regardless of injecting time. But, there was no difference in the response between the treated with small dose of MCA (0.5mg). Whereas erythrocyte-antibody(EA) rosette or erythrocyte-antibody-complement(EAC) rosette forming cells were significantly depressed by MCA. Under small dose of MCA (0.5mg), any difference of NK activity was not observed in all course of injecting time. But, under large dose of MCA, the activity was markedly inhibited to about half the values seen in control and this suppression was transient, resulting that the normal level was reached again 19 days after MCA. These results, which conform with the predictions of immunosuppression hypothesis, suggest that MCA inhibits immunological responses including NK activity and thereby allows the outgrowth of antigenic neoplastic cells.

• 대한의생명과학회지
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• 제16권4호
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• pp.365-376
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• 2010

The purpose of this study was to clarify the effect of light emitting diode (LED) irradiation on healing of impaired wound and alteration of mast cells in experimental diabetic rats. Twenty-four male Sprague-Dawley rats were divided into four groups: excision (Ex), excision-LED irradiation (Ex-LED), diabetes + excision (DM) and diabetes + excision + LED irradiation (DM-LED). Diabetes was induced in rats by streptozotocin (STZ) injection (70 mg/kg, single dose) and 6 mm punch excision wounds were created on the back after shaving hair. The LED-irradiated rats were treated to a daily dose of $5\;J/cm^2$ LED (630 nm) light for 11 days after surgery, and were killed at day 1, 3, 7 and 11. The lesion and adjacent skin tissues were excised, fixed with 10% buffered formalin and embedded with paraffin. For evaluation of wound healing, hematoxylin-eosin (HE) and Masson trichrome staining were performed. Mast cells (MCs) were stained with toluidine blue (pH 0.5) and quantified using a computerized image analysis system. The proliferation activity of keratinocyte in skin tissues was analyzed on sections immunostained with proliferative cell nuclear antigen (PCNA). The results showed that wound healing rate, collagen density and neo-epidermis length, number of PCNA-positive cells, fibroblasts and mast cells were significantly higher in the LED-irradiated rats than in the DM and Ex rats throughout the periods of experiment. Exceptionally, the number of MCs was significantly lower at day 11 compared with day 7 after surgery in the all groups. These findings suggest that the LED irradiation may promote the tissue repair process by accelerating keratinocyte and fibroblast proliferation and collagen production in normal rats as well as in diabetic rats, and MCs may play an important role at an early stage of skin wound healing in normal and diabetic rats.