• 대한한방부인과학회지
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• 제19권2호
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• pp.77-91
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• 2006

Purpose : To address the ability of Dohaekseungkitang (DST: a commonly used herb formulation in Korea, Japan and China to have anti-cancer effect on cervical carcinoma), we investigated the effects of DST on programmed cell death (apoptosis) in HeLa human cervical carcinoma cells. Methods : We cultured HeLa cell which is human metrocarcinoma cell in D-MEM included 10% fetal bovine serum(Hyclone Laboratories) below $37^{\circ}C$, 5% CO2. Then we observed apoptosis of log phage cell which is changed cultivation liquid 24 Hours periodically. Results : After the treatment of DST for 48 hours, apoptosis occurred in a dose-dependent manner. In this study, we have shown that DST induces calpain and the associated caspase-8 and -9 activations. Apoptosis was prevented by pre-incubation of the cells with the calcium cHeLator-BAPTA-AM, calcium channel blocker-Nif edipine or Ryonidine agonist-Ryonidine peptide, implicating calcium in the apoptotic process. Ubiquitous calpains (mu- and m-calpain) have been repeatedly implicated in apoptosis, especially in calcium-related apoptosis. However this study showed 1hat either calpain inhibitor-calpastin or caspase-3 inhibitor-DEVD- did not blocked the herb formulation-induced apoptosis in HeLa human cervical carcinoma cells. D ST initiates a cell death pathway that is partially dependent of caspases. DST-induced apoptosis requires caspase-independent mechanism. Conclusion : We conclude that DST-induced calpain activation triggers the intrinsic apoptotic pathway in which caspase-independent mechanism is also involved.

• 대한한방부인과학회지
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• 제20권1호
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• pp.32-49
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• 2007

Purpose: Breast cancer is the most common disease in Korean women. Despite remarkable improvements in treatment strategies against various cancer during the past 40 years, breast cancer still remains as one of the main causes of cancer mortality among women in the whole world. This study was carried out to investigate antioxidative and anti-proliferative effects on MCF-7 human breast cancer cells of Ikiyangyoung-Tang extract. Methods: We measured a content of polyphenol and flavonoid in the Ikiyangyoung-Tang extract, eliminative ability of DPPH radical, ABTS free radical and hydrogen peroxide, antioxidative effects of linoleic acid, cytotoxicity on MCF-7 human breast cancer cells. MCF-7 cells were cultured in Dulbecco's modified Eagle's medium/F12(DMEM/F12) supplemented with 10 % fetal bovine serum(FBS; Gibco) and antibiotics. Results : The extract of Ikiyangyoung-Tang contains polyphenol of 168.3${\pm}$12.8 ${\mu}$g/mg and flavonoid of 84.3${\pm}$3.4 ${\mu}$g/mg. Above results show profitable abilities of elimination of ${\alpha}$-${\alpha}$-Diphenyl-${\beta}$-picrylhydrazyl(DPPH) radical, ABTS free radical and hydrogen peroxide. Also, the extract of Ikiyangyoung-Tang strongly inhibits the proliferation of MCF-7 cells in a dose ependent manner. And. it has cytotoxicity on NIH3T3 cells. Conclusion : It can be concluded that Ikiyangyoung-Tang extract has an antioxidative effect and antiproliferative effect on MCF-7 human breast cancer cells.

• Journal of Periodontal and Implant Science
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• 제32권1호
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• pp.235-247
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• 2002

The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of human gingival fibroblasts. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells were inoculated in the multiwell plates coated with chitosan in concentration of 0.02, 0.2, and 2 mg/ml. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized nodules was evaluated after 21 days of culture. The results were as follows : The morphology of cells on the chitosan-coated well was round or spheric. Round cells were aggregated since 6 hours of culture and showed nodule-like appearance after 24 hours of culture and did not achieved confluency at 7 days. The attachment of gingival fibroblasts was inhibited by chitosan coating with a tendency of dose dependent pattern. But, cellular activity of unit cell was higher than control. The proliferation of gingival fibroblasts was inhibited by chitosan coating at 2 mg/ml(P<0.01), while the cell proliferation at 0.02, 0.2 $mg/m{\ell}$ was comparable to the control well. Total alkaline phosphatase activity was inhibited by chitosan coating and decreased in the course of time. While ALP activity of unit cell was the highest at 2mg/ml after 4 days of culture. Finally, gingival fibroblasts produced the mineralized nodule at 2 mg/ml. In summary, the attachment, proliferation, and alkaline phosphatase activity of gingival fibroblasts were influenced differently by the concentration of coated chitosan. From this study, it could be used as the matrix of tissue engineering for gingiva without inhibition on proliferation of gingival fibroblasts using chitosan at the optimal concentration (0.02mg/ml).

• Journal of Preventive Medicine and Public Health
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• 제22권4호
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• pp.518-527
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• 1989

The relationship between copper content in scalp hair and mental retardation was investigated. Samples of scalp hair were collected from 297 mentally retarded children who were students in one of two schools providing special educational services, one, consisted of children living in an orphan home, the other, children living with parents. For comparison, 117 scalp hair samples were collected from the children who had got average or above average academic achivement in a regular elementary school. Hair samples were taken from the nape of the neck and the copper content was determined by an atomic absorption spectrophotometer (IL 551). There was no statistically significant difference in scalp copper levels across different age groups except female orphan group, but no trend or correlation between copper conents and age was found. The hair copper contents of the mentally retarded children groups were significantly lower than that of control groups. But there was no dose-response relationship between degree of mental retardation and hair copper level. The hair copper contents of the group accompanied by Down's syndrome and unknown group were significantly lower than that of control group in both sex, and in the case of accompanied by epilepsy or autism, lower than control group in male. Although the results of this study show no evidence that mental retardation has owed to copper deficiency, the possibility of copper deficiciency in their fetal or infant age could not be ruled out. Thus further study is needed to determine whether mental retardation could be attributed to copper deficiency, through the examinations about their living environments, dietary pattern, eating habit and the impact of copper deficiency on brain development.

• 대한약침학회지
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• 제23권4호
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• pp.212-219
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• 2020

Objectives: Nowadays cancer treatment is an important challenge in the medical world that needs better therapies. Many active secretions produced by insects such as honey bees used to discover new anticancer drugs. Bee venom (BV) has a potent anti inflammatory, anti cancer and tumor effects. The aim of present study is evaluation of anticancer effects induced by Apis mellifera venom (AmV) on cell Lines. Methods: AmV was selected for study on cancer cell lines. Total protein, molecular weight and LD50 of crude venom were determined. Then, cells were grown in Dulbecco's Modified Eagle medium supplemented with 10% fetal bovine serum and 1% antibiotics. The A549, HeLa and MDA-MB-231 cell Lines were exposed by different concentration of AmV. The morphology of cells was determined and cell viability was studed by MTT assay. Evaluation of cell death was determined by and DNA fragmentation. Results: The results from MTT assay showed that 3.125 ㎍/mL of A549, 12.5 for HeLa and 6.25 ㎍/mL of MDA-MB-231 killed 50% of cells (p < 0.05). Morphological analysis and the results from hoescht staining and DNA fragmentation indicated that cell death induced by AmV was significantly apoptosis. Conclusion: The data showed that using lower dosage of AmV during treatment period cause inhibition of proliferation in time and dose dependant manner. Findings indicated that some ingredients of AmV have anticancer effects and with further investigation it can be used in production of anticancer drugs.

• 대한한방부인과학회지
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• 제15권4호
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• pp.45-60
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• 2002

생쥐의 calvarial osteoblast세포를 분리배양하여 gelatinase생성여부를 골흡수과정에서의 역할을 규명하기 위하여 SDS-PAGE-zymography분석을 한 결과 progelatinase-A를 항속적으로 합성하고 있음을 확인하였다. 생쥐의 osteoblasts를 골재흡수 약물인 PTH, $1,25(OH)_2D_3$, 단핵구배양액 (MCM) 그리고 IL-1으로 자극시키면 gelatinase생산을 촉진하여 콜라겐분해가 증가되었으나, indomethacin과 dexamethasone은 생쥐의 osteoblastic세포의 collagenolysis를 저해하였다. 한편, 골재흡수에 IL-1을 생쥐태아 유래의 장골조직배양 (fetal mouse long bone organ culture)에 처리하자 IL-1 은 골재흡수를 촉진하였다. 더우기, $IL-1{\alpha}$의 농도의존성에 대한 indomethacin과 dexametasone의 영향을 검토한 결과 직선형의 비례커브로 영향을 미쳤다. 이러한 골대사의 지견을 바탕으로 대영전-자하거의 열수추출물의 시험관내 독성검사에서 $1-200\;{\mu}g/ml$의 농도에서는 독성이 없었으며, 또한, $300\;{\mu}g/ml$ 농도에서도 생쥐의 calvarial골에는 독성이 없었다. 대영전-자하거 extract는 PTH (2 units/ml), MCM (5%, v/v), $rhIL-1{\alpha}$ (1 ng/ml) $1,25(OH)_2D_3$ (10 ng/ml)처리에 대해서 그리고 $IL-1{\alpha}$와 $IL-1{\beta}$-유발 collagenolysis에 대해서도 보호효과가 있었다. 대영전-자하거extract을 1시간동안 전처리와 후처리에서 콜라겐분해에 약간의 보호활성이 있었으며 $IL-1{\alpha}$와 $IL-1{\beta}$에 의해 유발되는 콜라겐분해에 보호활성이 보였다. 1시간동안 전처리는 콜라겐분해를 감소시키며, 대영전-자하거 extract는 gelatinase효소를 저해하였으며 PTH, $1,25(OH)_2D_3$, $IL-1{\beta}$ 및 $IL-1{\alpha}$로 유발된 효소활성화가 저해되었다. 즉, 대영전-자하거 extracts는 $IL-1{\alpha}-$와 $IL-1{\beta}$에 의해 촉진되는 골재흡수에 효과적이었으며, 비스테로이드성 항염증제제 (indomethacin 과 dexamethasone)에 의한 골재흡수방지 효과와 유사하였다. 이러한 결과는 대영전-자하거extract가 골다공증치료에 효과적임을 나타내는 것이다.

• Journal of Periodontal and Implant Science
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• 제26권2호
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• pp.448-468
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• 1996

To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.

• The Korean Journal of Pain
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• 제14권1호
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• pp.61-67
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• 2001

Background: Usually, lumbar epidural block is performed on the $L_{3-4}$ interspace. This study was designed to evaluate the analgesic efficacy and shortening of labor duration comparing the $L_{1-2}$ and $L_{3-4}$ interspace epidural blocks in nulliparous normal vaginal deliveries and then investigates side effects following the blocks. Methods: Eighty healthy nulliparous women were divided into two groups, $L_{1-2}$ (n = 40) and $L_{3-4}$ (n = 40). Epidural blocks, lumbar epidural block were performed at the $L_{1-2}$ and $L_{3-4}$ interspace with a catheter advancing 3 cm cephalad. The initial dose of 12 ml (0.167% bupivacaine, fentanyl $50{\mu}g$ and clonidine $75{\mu}g$) was injected epidurally at 4 cm dilatation of cervix and severe pain of labor. If a visual analogue scale (VAS) score was more than 4 points, an additional dose was administered epidurally using the same volume as the above mentioned, but with the exception that the bupivacaine was diluted to 0.1 percentage. The maternal blood pressure, pulse rate, respiration rate and fetal heart rate were measured at 10 min intervals for the first 30 min, at 15 min interval for the next 30 min and at 30 min interval for the last one hour following the blocks. The duration of the first (active) and second stages of labor was counted and the neonatal Apgar score was recorded at one and five min after delivery. The degree of motor block, pruritus, nausea and vomiting were also noted. Results: The patients in group $L_{1-2}$ had lower pain scores than group $L_{3-4}$ at 5, 20, 30, 60 mins. The duration of 1st and 2nd labor stage in the $L_{3-4}$ epidural block were $272{\pm}33.5$ min, $49.2{\pm}27.4$ min respectively but those in the $L_{1-2}$ epidural block were $253.5{\pm}32.5$ min, $37.3{\pm}22.3$ min, respectively. Conclusions: We concluded the analgesic efficacy and shortening of labor duration in $L_{1-2}$ epidural block was better than those in $L_{3-4}$ epidural block. Maternal hemodynamic change, motor block. pruritus, nausea, vomiting and Apgar score showed no significant differences between the two groups.

• Journal of Periodontal and Implant Science
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• 제27권4호
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• pp.685-700
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• 1997

The ultimate aim of periodontal treatment is periodontal regeneration, which necessiates the regeneration of bone tissues. This paper investigated the effect of growth factor on bone cells. Platelet-derived growth factor(PDGF) is the one of the polypeptide growth factor that has been reported as a biological mediator which regulates activities of the cell proliferation, migration and metabolism of undifferentiated mesenchymal cells. The purpose of this study is to evaluate the effects of PDGF on bone nodule formation and ALP activity of MC3T3-El cells. Cells were seeded at $1{\times}10^5cells/well$ in alpha-modified eagle medium containing 10% fetal bovine serum, lOml beta-glycerophosphate and $50{\mu}g/ml$ of ascorbic acid. PDGF 0, 0.1, 1, 10 ng/ml were added to the cells at a confluent state and cultured for 3, 7, 14, 21, 28 days. We examined bone nodule formation and alkaline phosphatase activity. The results were as follows : There were bone nodule formation at day 21 both in control and all the experimental groups, and at day 28, all the experimental groups showed much more bone nodules than control groups. Compared to control-l group, ALP activity was increased in PDGF O.1ng/ml group and was decreased in 1,10ng/ml PDGF treated groups.{P< 0.05, P< 0.01) Compared to control-2, ALP activity was decreased in all the experimental groups except PDGF 0.1ng/ml in 21 day group. In the time-response effect, ALP activity was increased by the day 14 in all the experimental groups and thereafter ALP activity was decreased.(P<0.05, P< 0.01) In the dose-response effect, ALP activity was decreased as the dose of PDGF was increased, and after 21 day ALP activity was lowest in 1 ng/ml group, ALP activity was highest in the day 7 in control group and 0.1 ng/ml, 14 day experimental group. In conclusion, PDGF is considered more effective in the proliferation than differentiation of osteoblast-like cells, and it may be useful to study the combined effect of PDGF and other growth factors on osteoblast-like cells.

• 대한수의학회지
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• 제34권1호
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• pp.165-172
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• 1994

DA-125는 새로운 안트라사이클린계 항암성 항생제로서 아드리아마이신의 유도체이다. Sprague-Dawley 랫트를 이용하여 DA-125의 배아 및 태자독성발현능력을 조사하였다. 교미확인(정충확인일=0일)된 120마리의 랫트를 4개군으로 나눈후 0, 0.1, 0.3 및 1.0mg/kg의 용량으로 임신 7일 부터 임신 17일까지 1일 1회 연속 정맥투여 하였으며 임신 20일째에 제왕절개를 하여 태자를 적출하였다. 1mg/kg 투여군에서는 모동물의 사료섭취량의 감소, 체중감소 및 비장중량의 감소와 배아 흡수율의 증가 및 태자체중의 감소가 관찰되었다. 또한 여러가지 종류의 외표, 내부장기 및 골격기형들이 각각 11.9, 41.8 및 14.5%의 빈도로 출현했다. 그중 특이 기형소견으로는 뇌탈출증, 복벽파열, 외측 및 제3뇌실의 확장, 늑골유착 등을 들 수 있다. 0.1 및 0.3mg/kg 투여군에서는 어떠한 배아 및 태자 독성증상도 나타나지 않았다. 이상의 결과에서 DA-125는 랫트에 있어서 경미한 모독성 용량에서 배아 및 태자독성효과를 나타냄을 알 수 있었다.