Animals ; Humans ; Lymph Nodes ; pathology ; Lymphangiogenesis ; physiology ; Lymphatic Metastasis ; Lymphatic Vessels ; metabolism ; pathology ; Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor C ; metabolism ; physiology ; Vascular Endothelial Growth Factor D ; metabolism ; physiology

To investigate the effect of acid and alkali stress on ginsenoside content of Panax ginseng, adventitious roots culture in bioreactors were incubated for 30 d and pH value was adjusted. Ginsenoside content increased by reducing or raising the pH in culture medium, the muxium ginsenoside content was determined on the 5th days after acid treatment and on the 7th days after alkali treatment. The result of histochemical localization of ginsenoside revealed that the red color from light to dark were found in the adventitious root tissue, and ginsenoside mainly located in the pericycle cells where appeared the dark red color.
Ginsenosides ; metabolism ; Hydrogen-Ion Concentration ; Panax ; metabolism ; physiology ; Plant Roots ; metabolism ; Stress, Physiological ; Time Factors

To improve cell suspension culture system of Panax ginseng, the dynamic of cell growth and medium consumption were studied, and the effects of filter on the culture vessel, revolution number, and inoculation density on cell growth and ginsenoside accumulation were also investigated. The maximum cell growth and ginsenoside accumulation was found on the 20th days of suspension culture, therefore, 20 days were confirmed as a suitable culture period for mass production of ginsenoside. Cell growth and ginsenoside content were promoted when the culture vessel had a ventilated filter. Revolution speed during suspension culture affected cell growth, but not ginsenoside content, a peak of ginsenoside productivity was found in the treatment of 120 r x min(-1). Inoculation density also influenced cell growth and ginsenoside accumulation, inoculation density of 6 g was better than other inoculation densities, the ginsenoside content and productivity were up to 12.8 mg x g(-1) DW and 146.6 mg x L(-1), respectively.
Cell Culture Techniques ; methods ; Cell Proliferation ; Culture Media ; chemistry ; Ginsenosides ; metabolism ; Panax ; cytology ; growth & development ; metabolism ; Suspensions

Objective: To investigate the effect of RNA interference (RNAi) targeting AKT1 and PI3K P85 on the proliferation and invasion of breast carcinoma MCF-7 cells. Methods: The recombinant adenovirus expression vector, which contained short hairpin RNA (shRNA) targeting open reading frames of AKT1 and PI3K P85 (rAd5-siAKT1-siPI3K), was transfected into human breast carcinoma MCF-7 cells. AKT1 and PI3K P85 mRNA and protein expressions were detected by real-time PCR and Western blotting analysis. The expressions of PCNA, cyclinD1, and P53 were also detected by Western blotting analysis. The proliferation and apoptosis of MCF-7 cells were measured by MTT, flow cytometry and 2-dementinal and 3-dementional matrigel assay. Results: Recombinant adenovirus vector rAd5-siAKT1-siPI3K dramatically down-regulated AKT1 and PI3K P85 mRNA and protein expressions in MCF-7 cells; the downstream factors PCNA and cyclin D1 were also down-regulated, while P53 was up-regulated. Growth of MCF-7 cells was inhibited by over 50% in rAd5-siAKT1-siPI3K group as measured by MTT assay, and cell cycle was arrested in G_1/G_0 phase compared with untransfected and rAd5-siCtrl transfected groups. Cell growth on matrigel matrix showed normal cell shapes, while the cells in rAd5-siAKT1-siPI3K transfected group were detached from the matrix or grew in scattered clustering patterns, forming only small aggregates. Conclusion: shRNA targeting AKT1 and PI3K P85 can significantly down-regulate the expression of AKT1 and PI3K P85 in breast carcinoma MCF-7 cells, and inhibit the growth of MCF-7 cells in vitro.

Objective: To study the effect of and possible mechanism of knockinng down PI3Kp85α using siRNA in MCF-7 human breast cancer cell line. Methods: Oligofectamine was used to transfect PI3Kp85α siRNA to knock down the PI3Kp85α expression level in MCF-7 human breast cancer cell line in vitro. Real-time PCR was conducted to detect the expression of PI3Kp85α. The effect of PI3Kp85αsiRNA on the growth of MCF-7 cells was measured by MTT. The cell cycle distribution and cell apoptosis were detected by cell flow cytometry. Protein expression was evaluated by immunofluorescence staining and Western blot. Results: The expression of PI3Kp85 α was knocked down with PI3Kp85α siRNA in MCF-7 cells. Cell growth was delayed in PI3Kp85αsiRNA-treated group. Conclusion: The suppressive effect of PI3Kp85αsiRNA on the growth of MCF-7 human breast cancer cell line is significant and PI3Kp85α could be a candidate for gene therapy for breast cancer.

Objective To investigate the effects of different standard cross sections and angles on the measurement accuracy of induced postnatal fetal long bones. Methods Fetal long bones (femori and humeri) in 30 cases with induced abortion were measured utilizing ultrasound from different angles and /or at different directions. The values measured from different sections and angles with vernier calipers were compared prenatally and postnatally. Results There was no apparent difference between the pre-induced abortion and those of the post-induced abortion. The results in the 30 cases showed that: (1) the values measured from anterior 90 degree, the long bone length would best match with the bare long bone length up to 96.7%, the match rate of other angles and/or directions was up to 80%; (2) no apparent statistical difference was between the length of left and right bone and no difference was found using 4 different directions and 3 different angles; (3)there was no difference between the left and right femuri and humeri.Conclusions Though the measured value from anterior 90 degree direction was the most accurate one, the statistical analtical results showed no difference among 12 values measured from 3 different angles and/or 4 different directions.

Objective： The current study was designed to assess the value of radiography on surgical margins of osteosarcoma. Methods： Tumor extension showed on X-ray plane film, CT and MRI was measured in 29 patients, and these data were analyzed statistically associated with pathological examination. Result： The width of tumor on X-ray plane film, CT and MRI was 42.7± 17.7 mm,70.1± 22.7 mm,73.3± 27.7 mm, respectively, and the length was 71.6± 24.5 mm,84.4± 30.3 mm, 123.6± 36.5 mm, respectively. As the result of histologic examination, the width of the tumor was 72.9± 26.1 mm, and the length was 119.8± 34.8 mm. The aggressive extension on MRI was similar to that on histologic exmination, and were more accurate than that on X-ray film and CT. Conclusion： It was reliable that the surgical margins were estimated on MRI.

Objective To investigate the role of NADPH oxidase in Herpes simplex virus 1 (HSV-1)-induced matrix metalloproteinase 9(MMP-9)expression in murine microglial BV2 cells.Methods BV2 cells induced by HSV-1 were divided into normal control group,HSV-1 infection group,and two apocynin treatment groups(in which apocynin was administered at 0.5 and 1.0 mmol/L after HSV-1 infection).MMP-9 gelatinolytic activity in the supematants of the cell cultures Was assessed by zymography.Semi-quantitative reverse transcription polymerase chain reaction(RT-PCR)was used to detect the mRNA expressions of NADPH oxidase subunit p47phox and matrix metalloproteinase-9 (MMP-9),and the intracellular reactive oxygen species(ROS)levels were measured by dihydroethidium staining(DHE).Results Compared to the normal control group,HSV-1 infection of the cells resulted in significantly up-regulated mRNA expression of NADPH oxidase subunit p47phox and MMP-9,and also in a 2-fold increase in the intracellular ROS level(P＜0.05).These changes were attenuated by the application of apocynin,but the mRNA expressions of p47phox and MMP-9 and ROS level still remained significantly higher than those in the normal control group(P＜0.05).Conclusion HSV-1 may induce MMP-9 activation through the generation of NADPH oxidase-dependent ROS in murine microglia.

OBJECTIVETo investigate the level of occupational exposure to 5-fluorouracil (5-Fu) in the pharmacy intravenous admixture service (PIVAS) of a hospital, and identify the sources of 5-Fu contamination.

METHODSThe 5-Fu concentrations in air, on the surface of different areas in PIVAS and personal protective equipments were detected using UV-vis spectrophotometry.

RESULTSThe 5-Fu in air could not be detected. The 5-Fu concentrations on five different surfaces of biological safety cabinets were (22.00 +/- 6.35), (13.99 +/- 2.46), (14.13 +/- 0.72), (7.25 +/- 1.19) and (9.87 +/- 1.23) ng/cm2, respectively, which were significantly higher than those [(3.14 +/- 0.04), (5.43 +/- 0.65), (2.26 +/- 0.17), (2.26 +/- 0.17) and (3.63 +/- 0.46) ng/cm2] of corresponding controls (P < 0.05 or P < 0.01). The 5-Fu concentrations of the floor under cabinets [(18.19 +/- 5.22) ng/cm2], the floor in front of cabinets [(10.25 +/- 2.57)ng/cm2], the office floor [(11.64 +/- 2.53) ng/cm2], the terrace floor [(99.89 +/- 14.06 ) ng/cm2], the floor beside trash can in dressing room [(24.54 +/- 0.23) ng/cm2] were significantly higher than those of control [(3.36 +/- 0.11 ) ng/cm2] (P < 0.05 or P < 0.01). The 5-Fu concentrations of the tables in preparation room [(7.22 +/- l.04) ng/cm2] and the tables in office [(11.81 +/- 1.18) ng/cm2] were significantly higher than those of control [(5.56 +/- 0.14) ng/cm2] (P < 0.05 or P < 0.01). The 5-Fu concentrations of the indoor handle in preparation room were significantly higher than those of controls (P < 0.05 or P < 0.01). 5-Fu concentrations on the surfaces of outdoor handle and floor beside door in preparation room were not significantly increased compared with controls (P > 0.05). The 5-Fu concentrations on the surfaces of infusion bags, transfer box, transfer trays were significantly higher than those of controls (P < 0.05). The differences of 5-Fu concentrations between outer and inner masks and controls were not significant (P > 0.05). The 5-Fu concentrations of gloves of preparing and checking staffs were significantly higher than those of controls (P < 0.05 or P < 0.01).

CONCLUSIONThe preparing and checking process of 5-Fu and the treatment of medical wastes are major sources of 5-Fu contamination.

Antineoplastic Agents ; analysis ; Drug Administration Routes ; Fluorouracil ; analysis ; Humans ; Occupational Exposure ; Pharmacy Service, Hospital

OBJECTIVETo study the effects of c9,t11-conjugated linoleic acid on the killing ability of macrophage to B16-MB cells in C57 mice and explore its possible mechanism.

METHODSThe five levels of CLA was designed as 0, 25, 50, 75, 100 micro mol/L. After macrophage was treated with CLA for 24 h, the killing ability of macrophage on B16-MB cells was evaluated by MTT, The expression of C57 mice macrophage cytokine IL-6, TNF-alpha and iNOS mRNA was detected by RT-PCR. The expression of Erk protein was examined by Western Blot assay.

RESULTSThe inhibitory effect of macrophage on tumor cell depend on the treatment of the increased c9,t11-CLA level, at the same time, the expression of IL-6, TNF-alpha and iNOS mRNA increased, the expression of Erk decreased with the elevating dose of CLA.

CONCLUSIONSc9,t11-CLA could increase the killing ability of macrophage in mice to B16-MB cells, and it was associated with induction of IL-6, TNF-alpha and iNOS mRNA expression. We speculate that antitumor ability of CLA may be associated with taking part in body immune regulation action, and the effects of CLA on the killing ability of murine macrophage to B16-MB cells was not associated with the MAPKErk pathway.

Animals ; Blotting, Western ; Cell Division ; Cell Line, Tumor ; Coculture Techniques ; Dose-Response Relationship, Drug ; Interleukin-6 ; genetics ; Linoleic Acids, Conjugated ; pharmacology ; Macrophages ; drug effects ; physiology ; Melanoma, Experimental ; pathology ; Mice ; Mice, Inbred C57BL ; Mitogen-Activated Protein Kinases ; metabolism ; Nitric Oxide Synthase ; genetics ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; genetics