Velvet antler is widely used as a traditional medicine, and numerous studies have demonstrated its tremendous nutritional and medicinal values including immunity-enhancing effects. This study aimed to investigate different deer velvet extracts (Sample 1: raw extract, Sample 2: dried extract, and Sample 3: freeze-dried extract) for proximate composition, uronic acid, sulfated glycosaminoglycan, sialic acid, collagen levels, and chemical components using ultra-performance liquid chromatography-quadrupole-time-of-light mass spectrometry. In addition, we evaluated the cytotoxic effect of the deer velvet extracts on BV2 microglia, HT22 hippocampal cells, HaCaT keratinocytes, and RAW264.7 macrophages using the cell viability MTT assay. Furthermore, we evaluated acute toxicity of the deer velvet extracts at different doses (0, 500, 1000, and 2000 mg/kg) administered orally to both male and female ICR mice for 14 d (five mice per group). After treatment, we evaluated general toxicity, survival rate, body weight changes, mortality, clinical signs, and necropsy findings in the experimental mice based on OECD guidelines. The results suggested that in vitro treatment with the evaluated extracts had no cytotoxic effect in HaCaT keratinocytes cells, whereas Sample-2 had a cytotoxic effect at 500 and 1000 μg/mL on HT22 hippocampal cells and RAW264.7 macrophages. Sample 3 was also cytotoxic at concentrations of 500 and 1000 μg/mL to RAW264.7 and BV2 microglial cells. However, the mice treated in vivo with the velvet extracts at doses of 500–2000 mg/kg BW showed no clinical signs, mortality, or necropsy findings, indicating that the LD50 is higher than this dosage. These findings indicate that there were no toxicological abnormalities connected with the deer velvet extract treatment in mice. However, further human and animal studies are needed before sufficient safety information is available to justify its use in humans.

In this paper, we investigate to determine quality characteristics, fatty acid composition and cytotoxic effect of extracts and fractions from whole Lycopus lucidus Turcz. roots. Additionally, we evaluated cytotoxic activity against the growth of human fibrosarcoma cells (HT-1080) and human gastric adenocarcinoma (AGS), human colon cancer cell (HT-29) lines using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Acetone+methylene chloride (A+M) and methanol (MeOH) extracts from L. lucidus Turcz. were obtained through solvent extraction. Then we further fractionated both extracts with n-hexane, 85% aq. MeOH, n-butanol (n-BuOH) and water. In fatty acid composition, L. lucidus Turcz. contained 33.2% of 18:1n-9 and 1.81% of 18:3n-3, respectively. The incorporation of treatment with A+M and MeOH extracts and n-hexane, 85% aq. MeOH, n-butanol (n-BuOH) and water fractions dose-dependently increased cytotoxicity against the growth of HT-1080 and AGS, HT-29 cancer cells (p<0.05). The A+M extract had a higher inhibitory effect on the growth of all cancer cells in comparison to MeOH extract. Among the fractions, the 85% aq. MeOH and n-hexane fractions showed a higher inhibitory effect after proliferating the three cancer cells. These results suggest that the 85% aq. MeOH and n-hexane fractions have a potential to inhibit the growth of human cancer cell lines.

Malignant gliomas and glioblastomas are the most common type of primary brain tumors. The treatment of malignant glioma involves surgery, radiation, and chemotherapy. These therapies have not been successful in curing malignant glioma and typically associated with dismal prognosis. Therefore, we can investigate thymidine kinase activity and cytotoxic effect after transfer suicide gene in U-251 glioma cells. We assessed expression patterns of green fluorescence protein(GFP) after infected with adenovirus in U-251 cells. After infection of HSV-tk in U-251 cells, we observed thymidine kinase activity with [3H]-penciclovir and cytotoxic effect by treated with ganciclovir. We could observe that expression level of GFP was increased according to infected concentrations in U-251 cells. GFP was not expressed in 1moi and 10moi, and slightly expressed in 30moi. Expression level of GFP was largely increased in 50moi and almost cells expressed GFP in 100moi. GFP expression has shown clear image in 100moi compared with other concentrations. We also investigated thymidine kinase activity using [3H]-penciclovir after infection of suicide gene HSV-tk into U-251 cells. Thymidine kinase activity increased in 10moi concentration compared with empty adenovirus infection. We could find that thymidine kinase activity was elevated proportional to HSV-tk infection amount in 30moi and 50moi. For evaluation of cellular cytotoxic effect of HSV-tk, we treated ganciclovir to U-251 cells and assessed cytotoxicity by using MTT assay. We could identifiy that cytotoicity appeared in very low concentration of HSV-tk compared with cancer cells originated with other organs. Cytotoxic effect was shown about 15% of U-251 cells of total cells in 5moi. By infection 10moi of HSV-tk, cytotoxic effect was intensively increased and about 60% of U-251 cells became extinct. About 70% cells exhibited cytotoxic effect in 30moi and more than 80% cells also appeared cytotoxic effect by infection of HSV-tk in 50moi, 100moi, and 200moi. Therefore, we could confirm to gene expression in U-251 cells was increased proportional to infected gene concentrations. Also we could find that thymidine kinase activity elevated with according to infected concentration and cellular cytotoxic effect was shown in very low concentration and higher cytotoxic effect also appeared by infection of suicide gene HSV-tk into U-251 glioma cells. These results suggest that gene therapy with suicide gene will be successful in curing brain tumors containing malignant glioma and glioblastoma.

Suicide gene transfer has been study extensively for therapies in various human diseases. We can evaluate cellular activity of thymidine kinase and cytotoxic effect in colon cancer cells after suicide gene transfer. We observed cellular expression of green fluorescence protein after transfer with adenovirus into colon adenocarcinoma HCT-15 cells. After transfer HSVtk, we also estimated thymidine kinase activity using [3H]-penciclovir and cellular cytotoxicity by MTT assay. After transfer green fluorescence protein into HCT-15 cells, we could observed fluorescence expression in 10 moi concentration. Expression level of green fluorescence protein markedly increased in 30 moi and most of HCT-15 cells expressed green fluorescence protein in 100 moi. By infection with HSVtk in HCT-15 cells and HT-29 cells, thymidine kinase activity in HCT-15 cells was about two fold higher than that HT-29 cells. Thymidine kinase activity at 1 moi concentration makes no difference with 0 moi in both cells. At 10 moi concentration, thymidine kinase activity increased about three fold compared with 1moi in HCT-15 cells, but not observed high increase in HT-29 cells. Thymidine kinase activity at 100 moi showed about three fold increase in HCT-15 cells and one and a half fold in HT-29 cells compared with 10 moi. By treatment of HSVtk at various mois and ganciclovir to HCT-15 cells, we could find that increased cytotoxic effect according to HSVtk concentration. Cellular cytotoxic effect was slightly appeared at 5 moi concentration and intensively increased at 30 moi concentration, dead colon cancer cells were reached about 30% of total colon cancer cells. Cellular cytotoxic effect was consistently increased until 50 moi, and about 50% of cells at 100 moi and less then 50% of HCT-15 cells at 200 moi were survived. Finally, we can identify that suicide gene transfer into HCT-15 cells is performed according to concentration of suicide gene and thymidine kinase activity also increase with HSVtk concentration in both HCT-15 cells and HT-29 cells. Additionally, we also find that suicide gene therapy by HSVtk with ganciclovir intensively increase cellular cytotoxicity in colon cancer cells. Therefore, our findings suggest that suicide gene therapy by HSVtk can affect cytotoxicy for colon cancer cells and eventually seems to influence therapeutic efficacy.

Resistance to the induction of apoptosis is a possible me- chanism by which tumor cells can survive anti-neoplastic treatments. Melanoma is notoriously resistant to anti-neop- lastic therapy. Previous studies have demonstrated focal adhesion kinase (FAK) overexpression in melanoma cell lines. Given its probable role in mediating resistance to apo- ptosis, many researchers have sought to determine whether the downregulation of FAK in melanoma cells would confer a greater sensitivity to anti-neoplastic agents. Genistein is a known inhibitor of protein-tyrosine kinase (PTK), which may attenuate the growth of cancer cells by inhibiting the PTK- mediated signaling pathway. This present study was under- taken to investigate the effect of genistein on the expression of FAK and cell cycle related proteins in the G361 me- lanoma cell line. Genistein was found to have a preferential cyto- toxic effect on G361 melanoma cells over HaCaT normal ke- ratinocytes. Genistein decreased the expression of 125 kDa phosphotyrosine kinase and the FAK protein in particular. Genistein treatment did not affect the expression of p53 in G361 cells in which p21 is upregulated. The expression of cy- clin B and cdc2 was downregulated by genistein treatment. Taken together, our data indicate that genistein induces the decreased proliferation of G361 melanoma cells via the in-hibition of FAK expression and regulation of cell cycle genes. This suggests that the use of genistein may be a via- ble approach to future melanoma treatments.

Nicotine(2~15mg/kg 2주간, 100mg/kg 회 투여)을 생후 4개월령 수컷 생쥐에 투여한 후, 고환에 미치는 광학현미경적 소견은 다음과 같다. 1. Nicotine 2mg/kg 투여군에서는 정세관과 정세관 주위의 Leydig 세포도 핵과 세포질이 뚜렷한 정상적인 소견을 나타내었으나, 5 mg/kg 투여군에서는 Leydig 세포의 핵과 세포질이 다소 비후되었으며, 염색정도가 약하였다. 2. 10 mg/kg 투여군은 대부분 정세관내의 정자발

식용 및 약용작물중에서 암세포에 대하여 생체세포독성을 유발하는 소재를 선발하기 위하여 한국인종양에서 유래한 대장암세포주(SNU-C4)와 위아세포주(SNU-1)에 대해 MTT검색법으로 50% 세포성장저해농도(IC50)를 측정한 결과는 다음과 같다. 1. 암세포에 대해 생체세포독성활성을 가지는 작물을 선발하기 위하여 17종의 작물을 대상으로 검색한 결과 보리, 옥수수, 오미자에서 세포독성효과가 230μg/ml이하의 농도로 유의한 효과를 나타내었다. 2. 품종간의 세포독성효과의 차이를 검색하기 위하여 세포독성효과를 나타낸 작물중에서는 보리와 옥수수를, 세포독성효과를 미약하게 나타내거나 나타내지 않은 작물중에서는 벼와 귀리를 대상으로 품종별로 검색한 결과, 세포독성의 유무는 1차 검색과 동일한 경향이 나타났으나, 벼에서는 품종에 따라 현저하게 세포독성의 차이를 나타내었으며, 특이할만한 점은 유색미인 흑도에서 다른 품종에 비하여 50% 세포성장저해농도가 유의하게 낮게 나타났다.