Investigation into perturbed nucleoside metabolism and cell cycle for elucidating the cytotoxicity effect of resveratrol on human lung adenocarcinoma epithelial cells / 中国天然药物

In an effort to understand the molecular events contributing to the cytotoxicity activity of resveratrol (RSV), we investigated its effects on human lung adenocarcinoma epithelial cell line A549 at different concentrations. Cellular nucleoside metabolic profiling was determined by an established liquid chromatography-mass spectrometry method in A549 cells. RSV resulted in significant decreases and imbalances of deoxyribonucleoside triphosphates (dNTPs) pools suppressing subsequent DNA synthesis. Meanwhile, RSV at high concentration caused significant cell cycle arrest at S phase, in which cells required the highest dNTPs supply than other phases for DNA replication. The inhibition of DNA synthesis thus blocked subsequent progression through S phase in A549 cells, which may partly contribute to the cytotoxicity effect of RSV. However, hydroxyurea (HU), an inhibitor of RNR activity, caused similar dNTPs perturbation but no S phase arrest, finally no cytotoxicity effect. Therefore, we believed that the dual effect of high concentration RSV, including S phase arrest and DNA synthesis inhibition, was required for its cytotoxicity effect on A549 cells. In summary, our results provided important clues to the molecular basis for the anticancer effect of RSV on epithelial cells.

Synthetic genomics: the art of design and synthesis / 生物工程学报

Benefited from the rapid development of high-throughput sequencing, genome editing, DNA synthesis and functional genomics, synthetic genomics gains the momentum in this century. The entire genomes of several viruses and one prokaryote have been chemically synthesized and applied to drive normal cellular processes. The first eukaryotic genome synthesis project (Sc2.0) is on-going and about half of the genome has been synthesized and functionally tested. The Human Genome Project-Write (HGP-Write) was proposed in 2016, which pushes the tide of synthetic genomics to a position we have never seen before. Technologies on genome-scale design and DNA synthesis have been rapidly developed, aiming to construct a more predictable and controllable genome at reasonable cost. The generation of synthetic organisms not only has promising applications for industry, environment, healthy and basic researches, but also raises ethic and policy concerns. This review presents the development of synthetic genomics, with emphasis on technologies for whole genome design, synthesis and assembly. We also discussed ethics, prospective and challenge in synthetic genomics. As one of the major branches in synthetic biology, synthetic genomics is still at its infant stage. A lot of excitement will come in the next few years.

La proliferación de los miocitos ventriculares del corazón de mamífero adulto: un fenómeno esporádico pero factible / Proliferation of adult mammalian ventricular cardiomyocytes: A sporadic but feasible phenomenon

La proliferación de los miocitos que forman parte de los ventrículos cardíacos del mamífero adulto ha sido descartada por algunos investigadores con el argumento de que estas células están diferenciadas en forma terminal; sin embargo, este dogma ha sido puesto en duda a partir de los hallazgos de otros investigadores quienes han observado que estos miocitos pueden presentar los procesos necesarios para la proliferación, es decir síntesis de ADN, mitosis y citocinesis, cuando el miocardio se daña en forma experimental con estrategias de tipo farmacológico o quirúrgico, o debido a condiciones patológicas relacionadas con el sistema cardiovascular. Esta revisión integra algunos de los trabajos disponibles en la literatura que han evaluado la síntesis del ADN, mitosis y citocinesis en estas células, en el miocardio dañado, para saber si su proliferación puede ser considerada como un fenómeno factible. La revisión concluye con una reflexión sobre las perspectivas del conocimiento generado en esta área de estudio.

Proliferation of adult mammalian ventricular cardiomyocytes has been ruled out by some researchers, who have argued that these cells are terminally differentiated; however, this dogma has been rejected because other researchers have reported that these cells can present the processes necessary to proliferate, that is, DNA synthesis, mitosis and cytokinesis when the heart is damaged experimentally through pharmacological and surgical strategies or due to pathological conditions concerning the cardiovascular system. This review integrates some of the available works in the literature evaluating the DNA synthesis, mitosis and cytokinesis in these myocytes, when the myocardium is damaged, with the purpose of knowing if their proliferation can be considered as a feasible phenomenon. The review is concluded with a reflection about the perspectives of the knowledge generated in this area.

Inhibition of DNA synthesis and radiosensization effects of thalidomide on esophageal carcinoma TE1 cells / 中华放射医学与防护杂志

Objective To explore the radiosensitization effect of thalidomide combined with X-ray on esophageal carcinoma TE1 cells.Methods Cell scratch assay Was used to detect the inhibition ability of different concentration of Thalidomide on cell invasion and metastasis.H3-TdR incorporation assay Was used to investigate the inhibition of DNA synthesis in TE1 cells by treated with Thalidomide singly or combination with X-rays.The colony formation assay Was used to analyze the radiosensitization of Thalidomide effect on TE1 cells.Results Thalidomide had obvious inhibition effect on TE1 cell metastasis.DNA synthesis and colony formation,which were correlated with drug concentration.The values D0,Dq and SF2 in TE1 cells were gradually decreased with thalidomide concentration increased.When the concentration of thalidomide was 100μg/ml,the SERD0 and SERDq were(1.4±0.2)and(1.5±0.1),respectively,While the concentration of thalidomide Was 1 50μg/ml,the SERD0 and SERDq were metastasis,DNA synthesis,and significantly enhance the radiosensitizing effect on esophageal carcinoma TE1 cells.

Effects of AG60 on Gastric Chief Cells of the Mouse Implanted with Ehrlich Carcinoma Cells / 대한해부학회지

This experiment was performed to evaluate the morphological responses of the gastric epithelial cells and the gastric chief cells of the mouse inoculated with Ehrlich carcinoma cells in the inguinal area following administration of acriflavine-guanosine composition (AG60). Healthy adult ICR mice were divided into normal and experimental groups. In the experimental groups, each mouse was inoculated with 1x10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. The day following the 7th injection of saline or AG60, each mouse was injected with methyl-3H-thymidine through tail vein. Seventy minutes after the thymidine injection, gastric tissues were taken and fixed in 10% buffered neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 and dried, and then placed in a light-tight box. The number of labeled epithelial cells in the gastric mucosae were observed and calculated. And for electron microscopic observation, gastric tissues were prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. The ultrathin sections were stained with uranyl acetate and lead citrate. The size of zymogen granules and mitochondria in the gastric chief cells were observed and calculated. On the autoradiographic study, number of labeled cells in the area of 3.5 mm width (6 micrometer thickness) of mouse gastric mucosae of normal control, tumor control and AG60-treated groups were 319.7+/-66.46, 343.7+/-47.72 and 102.3+/-54.99 respectively. On the electron microscopic study, the size of zymogen granule in the gastric chief cells of normal control, tumor control and AG60-treated groups were 0.74+/-0.208 micrometer, 1.18+/-0.291 micrometer and 0.97+/-0.259 micrometer, respectively. And the mitochondrial size of the gastric chief cells of normal control, tumor control and AG60-treated groups were 0.86+/-0.364 micrometer, 1.02+/-0.466 micrometer and 0.92+/-0.390 micrometer, respectively. And in the AG60 treated group, most chief cells did not show any difference in ultrastructure, except that myelin figures were more frequently observed, in comparison with that of nornmal control group. From the above results, AG60 may suppress the DNA synthesis of the gastric epithelial cells, but does not results severe fine structural defect on the gastric chief cells. These results suggest that AG60 is expected as one of the most effective anticancer drugs.

Analysis of DNA polymerase activity in vitro using non-radioactive primer extension assay in an automated DNA sequencer

Although different DNA polymerases have distinct functions and substrate affinities, their general mechanism of action is similar. Thus, they can all be studied using the same technical principle, the primer extension assay employing radioactive tags. Even though fluorescence has been used routinely for many years for DNA sequencing, it has not been used in the in vitro primer extension assay. The use of fluorescence labels has obvious advantages over radioactivity, including safety, speed and ease of manipulation. In the present study, we demonstrated the potential of non-radioactive in vitro primer extension for DNA polymerase studies. By using an M13 tag in the substrate, we can use the same fluorescent M13 primer to study different substrate sequences. This technique allows quantification of the DNA polymerase activity of the Klenow fragment using different templates and under different conditions with similar sensitivity to the radioactive assay.

Inhibitory effect of Poly gala tenyi folia willd on unscheduled DNA synthesis induced by Pb (CH_3COO)_2 in spermatogod of mice / 吉林大学学报(医学版)

0. 05) while UDS was induced by Polygala tenyi folia willd with various doses comparing with normal control. Nevertheless, Polygala tenyi folia willd in doses ranged from 1. 0 to 4. 0 g ? kg-1 inhibited strikingly UDS induced by Pb (CH3COO)2 (P

Effects of Antitumor Agents to the DNA Synthesis of Cecal Mucosa of Mouse / 체질인류학회지

This experiment was performed to evaluate the morphological responses of the cecal mucosa of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of 5-fluorouracil, mitomycin C or adriamycin. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups. In the experimental groups, each mouse was inoculated with 1 x 10(7) Ehrlich carcinoma cells subcutaneous in the inguinal area. From next day, 0.2 mL of saline, 5-fluorouracil (30 mg/kg), mitomycin C (400 microgram/kg) or adriamycin (2 mg/kg) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of anticancer drugs, each mouse was injected with a single dose of 0.7 micro Ci/gm of methyl-3H-thymidine (25Ci/mmol, Amersham Lab, England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed. The number of the labeled epithelial cells of the cecal crypts (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and evaluated. On histological study, in the experimental control and mitomycin C-treated mice, general morphology of the cecal mucosae was similar. And in the 5-fluorouracil-treated mice, slightly swelled epithelial cells and expanded lumen of the intestinal crypts were observed. But in the adriamycin-treated groups, slightly disrupted intestinal crypts, a large number of basophilic epithelial cells and the expanded lumen of the intestinal crypts were observed. On autoradiographic study, number of the labeled cells of normal control, experimental control, 5-fluorouracil treated, mitomycin C-treated, or adriamycin-treated groups were 362.2+/-56.12, 350.7+/-71.13, 215.7+/-80.55, 144.2+/-34.60 and 125.0+/-37.45, respectively. In the adriamycin and mitomycin C-treated groups, poorly-labeled cells containing only a few silver grains were observed more frequently than in those of the normal and experimental control groups. From the above results, adriamycin and mitomycin C suppressed the DNA synthesis of the epithelial cells of the cecal mucosa more severely as compared with 5-fluorouracil did. Especially, adriamycin was more harmful than mitomycin C and 5-fluorouracil on the cecal mucosae.

Effects of Antitumor Agents to DNA Synthesis in the Gastric Epithelia of Mice Implanted with Ehrlich Carcinoma Cells: An Autoradiographic Study / 대한해부학회지

This experiment was performed to evaluate the morphological responses of the gastric epithelium of the mouse, inoculated with Ehrlich carcinoma cells, following administration of 5-fluorouracil, adriamycin or mitomycin C. Healthy adult ICR mice weighing 25 gm each were divided into normal control and experimental groups (tumor control group, 5-fluorouracil treated group, adriamycin treated group, and mitomycin C treated group). In the experimental groups, each mouse was inoculated with 1 x 10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline, 5-fluorouracil (30 mg/kg), adriamycin (2 mg/kg) or mitomycin C (400 microgram/kg) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of anticancer drugs, each mouse was injected with a single dose of 0.7 microCi/gm of methyl-3H-thymidine (25Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed. The number of labeled epithelial cells in the gastric mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On histological study, in the gastric mucosae of adriamycin-treated groups, denatured surface epithelial cells, expanded lumen of the gastric gland, and congested lamina propria were observed. But in the 5-fluorouracil or mitomycin treated groups, severe morphological changes of the gastric mucosae were not observed. On autoradiographic study, numbers of the labeled cells in the gastric mucosae per 3.5 mm length of normal control, tumor control, 5-fluorouracil-treated, adriamycin-treated and mitomycin C treated groups were 267.3 (+/-48.86), 273.6 (+/-59.41), 375.3 (+/-83.36), 15.3 (+/-9.66) and 124.0 (+/-32.66), respectively. In the adriamycin and mitomycin C-treated group, poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently as compared in those of the normal control group. But in the 5-fluorouracil-treated group, number of the heavy labeled cells were observed more frequently than in those of the normal control group. From the above results, adriamycin and mitomycin C may severely suppress the DNA synthesis of the epithelial cells of the gastric mucosae. But some amount of the 5-fluorouracil (30 mg/kg) may not suppress the DNA synthesis of gastric epithelial cells.

Effects of Adriamycin or CP-2 to the Rectal Mucosae of Mouse Implanted with Ehrlich Carcinoma Cells: An Autoradiographic Study / 대한해부학회지

This experiment was performed to evaluate the morphological responses of the rectal intestinal glands of the mouse, inoculated with Ehrlich carcinoma cells, following administration of adriamycin or composition of the extracts of the Croton tiglium and Coptis chinensis rhizome (CP-2, Institute of Experimental Tumor Research, Seoul, Korea). Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (tumor control group, adriamycin treated group, and CP-2 treated group). In the experimental groups, each mouse was inoculated with 1 x 10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 mL of saline, adriamycin (2 mg/kg) or CP-2 (30 mg/kg) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of anticancer drugs, each mouse was injected with a single dose of 0.7 microCi/gm of methyl- 3H-thymidine through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed. and rectal tissues were collected and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab., England) in the dark room and dried. The number of the labeled epithelial cells of the rectal crypts (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On histological study, in the rectum of adriamycin treated groups, length of the intestinal crypts is shorter than those of the normal control ones. Disrupted intestinal crypts and epithelial cells were observed. But in the CP-2 treated group, morphological changes of the rectum were not observed. On autoradiographic study, number of the labeled cells of normal control, rumor control, adriamycin-treated, CP-2-treated groups were 263.1 (+/-38.65), 395.7 (+/-52.52), 73.3 (+/-22.54), 96.3 (+/-28.36), respectively. In the adriamycin and CP-2 treated groups., poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently than in those of the normal and tumor control groups. But in the tumor control group, number of the heavy labeled cells were observed more frequently than in those of the normal control group. From the above results, adriamycin and CP-2 may suppress the DNA synthesis of the cells of the rectal crypts. But CP-2 does not result any histological defect on the rectal mucosa. These results suggest that CP-2 is expected as one of most effective anticancer drugs.

Effects of 5-Fluorouracil, Mitomycin C or AG60 to the DNA Synthesis of Rectal Epithelium of Mice Implanted with Ehrlich Carcinoma Cells / 체질인류학회지

This experiment was performed to evaluate the morphological responses of the intestinal gland of the mouse, rectum inoculated with Ehrlich carcinoma cells, following administration of 5- fluorouracil, mitomycin C or AG60. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (tumor control group, 5-fluorouracil, mitomycin C treated group, and AG60 treated group). In the experimental groups, each mouse was inoculated with 1*10 (7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 mL of saline, 5-fluorouracil (30 mg/kg), mitomycin C (400 microgram/kg) or AG60 (5 mg/kg) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of anticancer drugs, each mouse was injected with a single dose of 0.7 microCi/gm of methyl-3H-thymidine through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed. The number of the labeled epithelial cells of the rectal crypts (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On histological study, in the rectum of mitomycin C treated groups, narrowed intestinal gland, a number of the nectotic changed epithelial nuclei and loosely arranged lamina propria were observed. But in the AG60 treated group, morphological changes of the rectum were not observed. On autoradiographic study, number of the labeled cells of normal control, tumor control, 5-fluorouracil (30 mg/kg) treated, mitomycin C (400 microgram/kg) treated and AG60 (5 mg/kg) treated groups were 246.3+/-42.30, 253.8+/-20.54, 172.7+/-19.02, 108.7+/-17.67 and 53.8+/-11.70, respectively. In the AG60 and mitomycin C treated group, poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently than in those of the normal control group. From the above results, AG60 (5 mg/kg) and mitomycin C (400 microgram/kg) are more suppressed the DNA synthesis of the cells of the rectal crypts as compare with 5- fluorouracil (30 mg/kg). And AG60 does not result any histological defect on the rectal mucosa. These results suggest that AG60 is expected as one of most effective anticancer drugs.

Effects of Antitumor Agents on the Duodenum of Mouse Implanted with Ehrlich Carcinoma Cells: An Autoradiographic Study / 대한해부학회지

This experiment was performed to evaluate the morphological responses of the intestinal gland of the mouse duodenum inoculated with Ehrlich carcinoma cells, following administration of 5-fluorouracil, mitomycin C or CP -2. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups. In the experimental groups, each mouse was inoculated with 1 x10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From the next day of inoculation, 0.2 mL of saline (experimental control group), 5-fluorouracil (30 mg/kg), mitomycin C (400 microgram/ kg) or CP -2 (30 mg/kg) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of anticancer drugs, each mouse was injected with a single dose of 0.7 microCi/g of methyl -(3)H-thymidine (25 Ci/mmol) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed. The number of the labeled epithelial cells of the duodenal crypts (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On histological study, in the duodenum of mitomycin C treated groups, narrowed intestinal gland, a number of the nectotic epithelial nuclei and loosely arranged lamina propria were observed. However, in the CP-2 treated group, morphological changes of the duodenum were not observed. On autoradiographic study, number of the labeled cells of normal control, experimental control, CP-2 treated, 5-fluorouracil treated and mitomycin C treated groups were 625.5 +/-58.85, 691.3 +/-82.32, 428.3 +/-83.16, 527.5 +/-79.84 and 297.33 +/-45.72, respectively. In the CP-2 and mitomycin C treated group, poorly-labeled cells containing only a few silver grains of (3)H-thymidine were observed more frequently than in those of the normal control group. From the above results, CP-2 and mitomycin C are more suppressed the DNA synthesis of the cells of the duodenal crypts as compare with 5-fluorouracil. But CP-2 does not result any histological defect on the duodenal mucosa. These results suggest that CP-2 is expected as one of most effective anticancer drugs.

Effects of annexin Ⅱ antisense vector on the growth of lung cancer cells / 第三军医大学学报

Objective To explore the effects of antisense vector of annexinⅡ gene on the growth of SPC A 1, a human lung cancer cell line. Methods The total RNA was isolated from human lung cancer cell line SPC A 1 and the target DNA fragments were amplified by RT PCR. The antisense expression vector was constructed by double restriction endonuclease cleavage directional clone method. Annexin Ⅱ antisense expression vector was introduced into SPC A 1 cells by liposome transfection reagent. The expression of annexin Ⅱ mRNA was analyzed by semi quantitative RT PCR. The effects of antisense vector of annexinⅡ gene on the growth of SPC A 1 were observed. Results The antisense vector of annexinⅡ gene was constructed and introduced into SPC A 1 cells successfully. Semi quantitative RT PCR showed that the annexin Ⅱ mRNA expression reduced by about two thirds in the transfected cells as compared with that in the untransfected cells. Compared with the untransfected cells, transfected cells decreased significantly in cell growth, clone formation efficiency in plating and DNA synthesis. Cell cycle was blocked in G 0 G 1 phase. Conclusion Annexin Ⅱ could promote the growth of lung cancer cells and may be helpful for the development of lung cancer.

The Effects of Recombinant Human BMP-7, Prepared from a COS-7 Expression System, on the Proliferation and Differentiation of Rat Newborn Calvarial Osteoblasts

A family of proteins, the bone morphogenetic proteins (BMPs), which promote osteoblast differentiation and bone mineralization, have recently been identified. One, BMP-7, has shown the ability to induce cartilage and bone formation processes. In this report, the possibility that other cell lines, to CHO cells, may also be available as host cells for the expression of hBMP-7 was validated. Recombinant human BMP (rhBMP) -7 was produced in COS-7 cells, as a processed mature disulfide-linked homodimer, with an apparent molecular weight of 36, 000. Examination of the expressions of the markers characteristic of osteoblast phenotypes showed that the rhBMP-7 specifically stimulated the inductions of alkaline phosphatase (ALP) (5-fold increase at 100 ng of rhBMP-7/ml), parathyroid hormone (PTH) -mediated intracellular cAMP production (4-fold increase at 100 ng of rhBMP-7/ml) and osteocalcin synthesis (5-fold increase at 100 ng of rhBMP-7/ml). In summary, the in vitro mineralization assay results provide evidence that the rhBMP-7 peptide, produced by COS-7 expression system, possesses intact biological activity. A similar pattern of biological activity was observed for the BMP-7 in COS-7 cells compared to the corresponding CHO cell expression system. Thus, these findings can be experimentally utilized for the production of rhBMPs for in vitro or in vivo studies.

Inhibition of indomethacin on proliferation and DNA synthesis of cultured hRPE cells / 眼科研究

ObjectiveTo investigate the inhibitory effects of indomethacin(IN) on proliferation and DNA synthesis of cultured human fetal retinal pigment epithelium(hRPE) cells in vitro.MethodsPrimary culture and subculture of hRPE cells were established in vitro first.Cultured hRPE cells were treated by various concentrations 50,100,200,400,600 μ mol/L(final concentration)of IN for 24h.After 24h,the amount of DNA in RPE cells was determined by the absorbance at 280nm of Nucleic Acid δ Protein Analysis.Cells proliferation of RPE were measured with methyl thiazolyl tetrazolium(MTT) assay method by adding 100,200,400,600,800,1000μ mol/L(final concentration) of IN for 12h.ResultsAfter added various concentrations of IN,the DNA concentrations were ( 101.1712± 15.5124),( 88.6400± 13.5845),( 72.3651± 7.7969),( 59.9089± 10.7229),( 51.2236± 8.7757)μg/ml respectively,P values were 0.000,0.000,0.000,0.000,0.000(q test) as compared to that ( 213.7351± 83.1572)μg/ml in 0μg/L IN.The A values added 100,200,400,600,800,1000μmol/L of IN were ( 0.2367± 0.0546),( 0.1687± 0.0695),( 0.0819± 0.03461),( 0.0656± 0.01759),( 0.0554± 0.02865),( 0.0508± 0.02775)respectively,P values were 0 .158,0.000,0.000,0.000,0.000,0.000(q test) as compared to ( 0.2674± 0. 04302) of A value of 0ug/L IN.ConclusionThe data suggested that IN can inhibit DNA synthesis and proliferation of hRPE cells in vitro in a dose dependent manner.

Effects of Adriamycin or AG60 to the DNA Synthesis of Duodenal Epithelium of Mice Implanted with Ehrlich Carcinoma Cells: An Autoradiographic Study / 체질인류학회지

This experiment was performed to evaluate the morphological responses of the intestinal gland of the mouse, duodenum inoculated with Ehrlich carcinoma cells, following administration of adriamycin or acriflavine -guanosine composition (AG60, Taerim Pharm. Co. Seoul, Korea). Healthy adult ICR mice weighing 25 g each were divided into normal and experimental groups (experimental control group, adriamycin treated group, and AG60 treated group). In the experimental groups, each mouse was inoculated with 1 x10 7 Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 ml of saline, adriamycin (2 mg/ kg), AG60 (5 mg/kg) or AG60 (30 mg/kg) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of anticancer drugs, each mouse was injected with a single dose of 0.7 microCi/gm of methyl -3 H -thymidine (25 Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed. The number of the labeled epithelial cells of the duodenal crypts (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On histological study, in the duodenum of adriamycin treated groups, vesiculated epithelial cells of the intestinal villi, expanded lumen of the intestinal gland (G) and loosely arranged lamina propria were observed. But in the AG60 treated group, morphological changes of the duodenum were not observed. On autoradiographic study, number of the labeled cells of normal control, experimental control, adriamycin -treated, AG60 (5 mg/kg)-, and AG60 (30 mg/kg)-treated groups were 595.3 +/-48.96, 715.+/-89.11, 96.0 +/-15.62, 632.0 +/-83.16 and 370.3 +/-49.65, respectively. In the adriamycin and AG60 30mg/kg -treated group, poorly -labeled cells containing only a few silver grains of 3 H -thymidine were observed more frequently than in those of the normal control group. But in the experimental control group, number of the heavy labeled cells were observed more frequently than in those of the normal control group. From the above results, adriamycin and AG60 (30 mg/kg) may suppress the DNA synthesis of the cells of the duodenal crypts. But AG60 does not result any histological defect on the duodenal mucosa. These results suggest that AG60 is expected as one of most effective anticancer drugs.

Effects of Ligustrazine Injection on DNA Synthesis of Vascular Endothelial Cell / 中国药房

OBJECTIVE:To study the effect of ligustrazine injection(LTZ)on DNA synthesis of vascular endothelial cell.METHODS:In vitro cultured cell line of human umbilical vein endothelial cells,ECV304,were adopted to measure the ef?fects of LTZ on DNA synthesis by 3 H-TdR incorporation.RESULTS:LTZ could inhibit DNA synthesis of ECV304in a dose-dependent manner.CONCLUSION:The mechanism of anti-angiogenesis of LTS might be associated with inhibiting DNA synthesis of vascular endothelial cell.

The effects of insulin-like growth factors on the proliferation of endometrial stromal cells from patients with and without endometriosis / 대한산부인과학회잡지

OBJECTIVES: To test the hypothesis that the endometrial stromal cells from patients with endometriosis responds differently to the insulin-like growth factor (IGF) compared with those from patients without endometriosis. METHODS: IGFs in peritoneal fluid (PF) from patients with endometriosis(n=18) and without endometriosis(n=12;control patients) were measured by radioimmunoassay. Endometrial stromal cells from patients with endometriosis and control patients were cultured in serum free media(SFM) in the presence or absence of PF or IGF-I(0.25-25 ng/ml) or IGF-II(5-50 ng/ml) and the proliferation of endometrial stromal cells were evaluated by [3H] thymidine incorporation test. All statistics were performed by ANOVA test and student's t-test. RESULTS: When added to SFM, IGF-I(1-25 ng/ml) increased thymidine incorporation in both endometrial stromal cells from patients with endometriosis and control patients in dose dependent manner and IGF-II(5-25 ng/ml) gave similar response in latter cells but not in former cells. Within low IGF-I(less than 100 ng/ml) PF group or high IGF-I(more than 100ng/ml) PF group, the type of endometrial stromal cells did not result in any difference in thymidine incorporation. However, regardless of the source of stromal cells, high IGF-I PF group produced a greater extent of thymidine incorporation than low IGF-I PF group in patients with endometriosis but not in control patients. Also, thymidine incorporation was higher in high IGF-I PF group of former patients than in the same group of latter patients. PF induced higher thymidine incorporation in endometrial stromal cells than the same levels(0.25-2.5 ng/ml) of IGF-I directly added to SFM. CONCLUSIONS: The effects of IGF-I in PF on endometrial stromal cells are similar regardless of their source and IGF-I is one of several growth factors that may participate in the growth of endometrial stromal cells in pelvic endometriosis.

ENHANCEMENT OF DNA SYNTHESIS IN CULTURED ADULT RAT HEPATOCYTES BY 5-HT THROUGH STIMULATION OF 5-HT2 RECEPTOR / 药物分析学报

Hepatocytes were isolated from livers of adult male Sprague-Dawley rats and cultured in Williams'E Medium with [3 H] thymidine. The effect of 5-hydroxytryptamine (5-HT) was investigated through adding various concentrations (10-8～10-3 mol/L) of 5-HT to the hepatocyte cultures in the presence or absence of epidermal growth factor (EGF) and insulin. The involvement of 5-HT2 receptor was examined by adding a 5-HT2 receptor antagonist, ketanserin (10-6 mol/L), to some of the cultures containing 5-HT. The increment of DNA synthesis was measured by [3 H] thymidine incorporation. The results showed that 5-HT2 (≥10-6 mol/L) significantly (P＜0.05) increased the amount of DNA synthesis induced by EGF and insulin in the cultured adult rat hepaptocytes. The effect of 5-HT in enhancing DNA synthesis began to appear at a concentration between 10-7 and 10-6 mol/L and reached maximum at concentrations of ≥10-4 mol/L. The enhancement of DNA synthesis by 5-HT was significantly (P＜0.05) antagonized by ketanserin, suggesting that this effect of 5-HT was mediated by 5-HT2 receptor subtype.

Biological Effects of bioactive glass and natural coral on periodontal ligament fibroblast-like cell behavior / 대한치주과학회지

The purpose of this study was to evaluate the effects of bioactive glass and natural coral on the human periodontal ligament fibroblast(HPLF) behaviors during the regeneration process of peridontium. To determine the cellular events occuring in the presence of the particles of bioactive glass and natural coral, HPLF were isolated from healthy premolar teeth extracted for orthodontic treatment. Cells were cultured in alphaMEM at 37degrees C, 5% CO2, 95% humidity incubator. Bioactive glass and natural coral were powdered, and each particled(<40micrometer) were placed on the cultured cells at the concentration of 0.3mg/ml, and l,0mg/ml for experimental group. In control group no particles were added. And each group was evaluated by examining the cell morphology under phase-contrast micrograph at 4 day and transmission electron micrograph(TEM) and scanning electron micrograph(SEM) at 14 day, alkaline phosphatase activity at 5 and 9 day, protain synthesis at 4 day, DNA synthesis at 1, 2, 3 and 4 day, cell proliferation at 1, 3, 5,7 and 9 day and the formation of bone nodule at 30 day after culturing all groups in mineralizing supplemented mediun. No significant changes in cell morphology by adding these two matirials were found under phase contrast microscopy and TEM, HPLF phagocytocized each particles suggesting that HPLF is involved in the process of resorbing each particles and that bioactive glass were more biocompatible than natural coral. The ALPase activity of bioactive glass 0.3 mg/ml was similar with control groups and all the rests of control groups were significantly low(P<0.01) indicating a transient dedifferentiation of HPLF in the presence of bioactive glass and natural coral particles. There were no significant differences of protein synthesis between all groups. The DNA synthesis in experimental groups were significantly lower than control groups at 1, 2 and 3 day (P<0.01) but became similar to control groups at 4 day. Between control groups, the DNA synthesis in bioactive glass 0.3mg/ml group was significantly higher than other groups(P (0.01). Cell proliferation in natural coral 1.0mg/ml and bioactive glass l.0mg/ml groups were significantly lower than control group at 3 day(P(0.05) and there were no differences at 5, 7, 9 day. There were more bone nodule formation in experimental groups than in control groups. In conclusion, these results indicated that bioactive glass and natural coral have some effects of a transient dedifferentiation on HPLF and regeneration of periodontal tissues, however any significant cytotoxic effect on HPLF by these two particles were not found.