• 동의생리병리학회지
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• 제25권5호
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• pp.849-856
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• 2011

This study was investigated the protective effect of Cultivated Wild Ginseng(WG) on the toxicities induced by cyclophosphamide(CP) in mice. Twenty-four male BALB/c mice were divided into non-treated normal group(n=6), CP treated control group(n=6), CP+WG treated CP+WG group(n=6), WG treated WG group(n=6). CP(100 mg/kg of b.w., i.p) was injected at 0, 7 th, 14 th, 21 th, and 28 th day of the experiment respectively. WG(4.4 g/kg, i.p.) was administrated for 35days. Body and organ(heart, liver, kidney, testis) weight were measured. Histopathological examination on the organ(heart, liver, kidney, testis), morphometric analysis, and BrdU immunohistochemistry on the testis were performed. Body weight was decreased following CP administration. In contrast, such a decrease was significantly attenuated by WG administration. CPK and AST of CP+WG group were significantly decreased compared with CP group. Histopathologically, cross sectional area of testis and diameter of seminiferous tubule were significantly increased in CP+WG group compared with CP group. BrdU labelled cells in the seminiferous tubules were remarkably decreased in CP group. Whereas the number of seminiferous tubules labelled with BrdU in spermatogonia was increased by CP+WG administration. The obtained results suggest that WG has protective effect on CP-induced toxicity. This effect might be mediated through the supplementation of vital energy.

• Archives of Plastic Surgery
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• 제32권4호
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• pp.533-538
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• 2005

Due to its safety and softness, autologous fat transplantation has been commonly performed for soft tissue correction. However, the injected fat is absorbed resulting in the reduction of volume of the graft by 40-60% within a few months. Thus, there was an attempt to use adipocytes differentiated from preadipocytes in vitro for transplantation. Differentiated adipocytes were biocompatible and matured with gradual volume increase at transplantation site in clinical study(unpublished data). In addition, they did not induce immune rejection in response to nonself lymphocytes in a mixed lymphocyte reaction(MLR)(unpublished data). The purpose of this study is to differentiate mouse preadipocytes following labeling into adipocytes to establish an animal model for allogenic transplantation. Preadipocytes isolated from inguinal and retroperitoneal fat pad of C57BL/6 mice were proliferated with growth medium by passage 3 and differentiated into adipocytes with different culture conditions after labeled with BrdU. At most suitable conditions, above 90% of preadipocytes were differentiated and BrdU labeling did not affect differentiation rate and function of differentiated adipocytes. These results demonstrate that BrdU-labeled adipocytes resulting from this in vitro differentiation protocol are useful for allogenic transplantation study.

• The Korean Journal of Physiology and Pharmacology
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• 제22권2호
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• pp.145-153
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• 2018

The subgranular zone (SGZ) of hippocampal dentate gyrus (HDG) is a primary site of adult neurogenesis. Toll-like receptors (TLRs), are involved in neural system development of Drosophila and innate immune response of mammals. TLR2 is expressed abundantly in neurogenic niches such as adult mammalian hippocampus. It regulates adult hippocampal neurogenesis. However, the role of TLR2 in adult neurogenesis is not well studied in global or focal cerebral ischemia. Therefore, this study aimed to investigate the role of TLR2 in adult neurogenesis after photochemically induced cerebral ischemia. At 7 days after photothrombotic ischemic injury, the number of bromodeoxyuridine (BrdU)-positive cells was increased in both TLR2 knock-out (KO) mice and wild-type (WT) mice. However, the increment rate of BrdU-positive cells was lower in TLR2 KO mice compared to that in WT mice. The number of doublecortin (DCX) and neuronal nuclei (NeuN)-positive cells in HDG was decreased after photothrombotic ischemia in TLR2 KO mice compared to that in WT mice. The survival rate of cells in HDG was decreased in TLR2 KO mice compared to that in WT mice. In contrast, the number of cleaved-caspase 3 (apoptotic marker) and the number of GFAP (glia marker)/BrdU double-positive cells in TLR2 KO mice were higher than that in WT mice. These results suggest that TLR2 can promote adult neurogenesis from neural stem cell of hippocampal dentate gyrus through increasing proliferation, differentiation, and survival from neural stem cells after ischemic injury of the brain.

• Journal of Korean Neurosurgical Society
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• 제38권2호
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• pp.121-125
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• 2005

Objective : Many recent reports have shown that the mature mammalian brain harbors multipotent stem cells, rendering the brain capable of generating new neurons and glia throughout life. Harvested stem cells from an adult rat are transplanted in order to evaluate the cell survival and differentiation. Methods : Using a percoll gradient with a high speed centrifugation method, we isolate neural stem/progenitor cells were isolated from the subventricular zone[SVZ] of a syngeneic adult Fisher 344 rats brain. For 14days expansion, the cultured cells comprised of a heterogeneous population with the majority of cells expressing nestin and/or GFAP. After expanding the SVZ cells in the presence of basic fibroblast growth factor-2, and transplanting then into the hippocampus of normal rats, the survival and differentiation of those cells were examined. For transplantation, the cultured cells were labeled with BrdU two days prior to use. In order to test their survival, the cells were transplanted into the dorsal hippocampus of normal adult Fisher 344 rats. Results : The preliminary data showed that at 7days after transplantation, BrdU+ transplanted cells were observed around the injection deposition sites. Immuno-fluorescent microscopy revealed that the cells co-expressed BrdU+ and neuronal marker ${\beta}$-tubulin III. Conclusion : The data demonstrate that the in vitro expanded SVZ cells can survive in a heterotypic environment and develop a neuronal phenotype in the neurogenic region. However more research will be needed to examine the longer survival time points and quantifying the differentiation in the transplanted cells in an injured brain environment.

• 동의신경정신과학회지
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• 제21권2호
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• pp.29-44
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• 2010

Objectives : The purpose of this study was to investigate the effect of Gentianae Radix on neurogenesis and apoptosis in ethanol- induced newborn rats hippocampus dentate gyrus. Methods : In vivo, laboratory animals were divided into three groups; Normal group(N), Control group(C) and Treated group (TG)(n=7 for each group). N were treated saline daily for five days. C were treated 1.5 g/kg ethanol and saline daily for five days. TG were treated 1.5 g/kg ethanol and 300 mg/kg Gentianae Radix daily for five days. BrdU(5-bromo-2-deoxyuridine) assay was used to test neurogenesis in the dentate gyrus. And TUNEL(Terminal deoxynucleotidyl transferase dUTP nick end labeling) assay was used to test apoptosis in the dentate gyrus. Three groups were measured body weight, serum ethanol concentration, BrdU-positive cells and TUNEL-positive cells in the dentate gyrus. In vitro, MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to test viability in SK-N-MC cells. BrdU assay was used to test neurogenesis in SK-N-MC cells. DNA fragmentation and caspase-3 enzyme activity assay were used to test apoptosis in SK-N-MC cells. And treated ethanol and Gentianae Radix of all in vitro tests were made various concentration. Results : In vivo, Gentianae Radix modulated ethanol-induced neurogenesis and apoptosis in newborn rats hippocampus dentate gyrus. In vitro, TG 100 ${\mu}g/ml$ have significantly modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells. And only TG 100 ${\mu}g/ml$ have significantly protected SK-N-MC cells from ethanol-induced cytotoxicity. Conclusions : Gentianae Radix may have the effect that modulated ethanol-induced neurogenesis and apoptosis in SK-N-MC cells.

• 동의생리병리학회지
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• 제23권5호
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• pp.1025-1034
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• 2009

This study was carried out to examine the protective effect of Cultured Wild Ginseng(CWG) on the acute and subacute toxicities induced by doxorubicin(Doxo) in mice. Heart and liver weight was decreased following Doxo administration. In contrast, such a decrease was significantly attenuated by CWG administration. The value of serum CPK in Doxo group was increased compared with normal group. But the value of CWG group were decreased significantly compared with the values of Doxo group in the liver of the Doxo group, cloudy swelling of hepatic cells and narrowing of sinusoids were observed. Whereas in the CWG group, well oriented hepatic cell cords and sinusoids were observed. In the testis of the Doxo group, necrotic and degenerative changes of the seminiferous tubules, especially beneath testicular membrane were observed. But those lesions were alleviated in CWG group. Cross sectional area of testis and diameter of semineferous tubule were significantly increased in CWG group compared with Doxo group. Body weight was reduced in Doxo group compared with normal group. In contrast, such a decrease was significantly attenuated by CWG administration atwa5th day. Spermatogenetic cells in seminiferous tubules were necrotic and desquamated and the cellularity of seminiferous epithelia was reduced in Doxo group. But those lesions were attenuated by CWG administration. Cross sectional area of testis and diameter of seminiferous tubule were significantly increased in CWG group compared with Doxo group. In addition, the increase in lipid peroxidation(LPO) in testis was inaddition, the, iout such a increased was significantly inhibited in CWG group. BrdU labelled cells in the seminiferous tubules were remarkably decreased in Doxo group. Whereas the number of seminiferous tubules labelled with BrdU in spermatogonia was increased by CWG administration. The obtained results suggest that CWG has protective effect on doxorubicin-induced toxicity. This effect might be mediated through the supplementation of vital energy.

• Korean Journal of Acupuncture
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• 제19권1호
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• pp.77-84
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• 2002

목적 : 소아의 정신적 및 신체적 발달지연에 사용되고 있는 신기환이 해마의 신경세포 생성에 미치는 영향을 관찰하고자 하였다. 방법 : 신기환이 세포생성에 미치는 영향을 알아보기 위하여 해마 세포인 HiB5에 신기환을 처리, 배양하여 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay를 통해서 세포생성을 관찰하였다. 또한 3주령의 Sprague-Dawley (S-D)계 흰쥐의 중완혈에 신기환 약침을 자침후 해마의 치상회에서 bromodeoxyuridine (BrdU) immunohistochemistry를 시행하여 세포생성을 관찰하였다. 결과 : HiB5 배양세포에서는 신기환을 처리시 배양시간과 농도별로 세포생성율이 증가하였다. 또한, 대조군의 치상회에서 BrdU 양성세포수는 $128.50{\pm}9.53$, 1 mg/kg 신기환 자침군은 $157.08{\pm}10.82$ 및 10 mg/kg 신기환 자침군은 $204.80{\pm}17.68$로 나타났다. 이러한 결과로 신기환은 어린 S-D계 흰쥐의 치상회에서 세포생성을 증가시키는 것으로 사려된다.

• Korean Journal of Acupuncture
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• 제27권2호
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• pp.49-56
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• 2010

목적 : 뇌의 신경세포 증식은 해마 치상회와 뇌실하영역에서만 나타나는 현상이다. Kainic acid(KA)를 이용한 간질 동물모델을 연구하던 중 침이 해마 치상회의 신경세포증식을 촉진하는 현상을 발견하여 이를 보고하고자 한다. 방법 : 수컷 ICR계 생쥐를 Saline(n=8), KA(n=8), KA+Acu(n=8)의 세 군으로 나누고, 모든 생쥐들에게 KA 주입 3일 전부터 1일 1회씩 5'-bromodeoxyuridine(BrdU)을 3일간 주입하였다. Saline군에는 멸균된 생리식염수를 뇌실 내에 주입하였고, KA군 및 KA+Acu군에는 $0.1{\mu}g$의 KA를 뇌실 내에 주입하였으며, KA+Acu군에 속한 쥐들에게는 KA 주입 2일전, 1일전, 주입 직후에 양쪽 소부(少府)(HT8)에 자침하였다. KA 주입 3시간 후 쥐의 뇌를 적출하고 해마 치상회부위의 BrdU 및 neuropeptide Y (NPY)의 발현을 측정하였다. 결과 : 소부(少府) 자침이 KA의 독성으로 인한 신경세포의 파괴를 줄여주었으며, BrdU 양성 세포 및 NPY를 유의하게 증가시켰다. KA 주입시 세포증식이 일어나긴 하나, 3시간 안에는 거의 일어나지 않는다. 결론 : 소부(少府) 자침이 해마 치상회의 신경세포증식을 촉진하며, 이는 KA의 효과가 아닌 KA 투여 전 소부(少府) 자침으로 인한 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제22권2호
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• pp.274-282
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• 2012

The object of this study was to assess the efficacy of Polycan from Aureobasidium pullulans SM-2001, which is composed mostly of beta-1,3-1,6-glucan, on osteoarthritis (OA)-induced by anterior cruciate ligament transection and partial medial meniscectomy (ACLT&PMM). Three different dosages of Polycan (85, 42.5, and 21.25 mg/kg) were orally administered once a day for 84 days to male rats a week after ACLT&PMM surgery. Changes in the circumference and maximum extension angle of each knee, and in cartilage histopathology were assessed using Mankin scores 12 weeks after Polycan administration. In addition, cartilage proliferation was evaluated using bromodeoxyuridine (BrdU). As the result of ACLT&PMM, classic OA was induced with increases in maximum extension angles, edematous knees changes, and capsule thickness, as well as decreases in chondrocyte proliferation, cartilages degenerative changes, and loss of articular cartilage. However, these changes (except for capsule thickness) were markedly inhibited in all Polycan- and diclofenac sodium-treated groups compared with OA control. Although diclofenac sodium did not influence BrdU uptake, BrdU-immunoreactive cells were increased with all dosages of Polycan, which means that Polycan treatment induced proliferation of chondrocytes in the surface articular cartilage of the tibia and femur. The results obtained in this study suggest that 84 days of continuous oral treatment of three different dosages of Polycan led to lesser degrees of articular stiffness and histological cartilage damage compared with OA controls 91 days after OA inducement, suggesting that the optimal Polycan dosage to treat OA is 42.5 mg/kg based on the present study.

• Toxicological Research
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• 제27권4호
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• pp.247-251
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• 2011

To clarify whether inhibitory effect of estrogen on liver tumor is associated with cell proliferation, we investigated its role in diethylnitrosamine (DEN)-induced rat preneoplastic lesions, with time sequenced manners. F344 male rats (n = 90) were divided into three groups at 5 weeks of age. The mini-osmotic pumps providing a continuous infusion of DEN was implanted into the abdominal cavity of each animal in group 1, 2 and 3 at 6 weeks of age. To see the effect of estrogen, pellet containing 1 or 10 ${\mu}g$ of estradiol-3-benzoate (EB) was implanted subcutaneously in the animals of groups 2 or 3, respectively, one week prior to DEN treatment. Ten animals of each group were euthanized at 10, 14 and 18 weeks after DEN treatment. Liver tissues at each time point were fixed in 10% phosphate-buffered formalin and were processed and embedded in paraffin and 5 ${\mu}g$ sections mounted on a silanized slide. Glutathione S-transferase placental form (GST-P) positive foci and 5-bromo-2-deoxyuridine (BrdU) labeling cells were detected at each time point. Area of GST-P positive foci in DEN+EB 1 or 10 ${\mu}g$ group was significantly decreased compared to DEN alone at 14 weeks (p < 0.01 or p < 0.05, respectively) an at 18 weeks (p < 0.05 or p < 0.01, respectively). BrdU index in DEN+EB 1 or 10 ${\mu}g$ groups was significantly decreased compared to DEN alone at 14 weeks and at 18 weeks (p < 0.01). Taken together, we conclude that EB treatment decrease the DEN-induced liver preneoplastic lesions and this may be associated with decrease of cellular proliferation.