RESUMEN
Objective:To investigate the killing effects in vitro of a new photosensitizer black titanium dioxide b-TP-700-mediated photodynamics on prostate cancer cell line PC-3 cells.Methods:The b-TP-700 was successfully prepared by using solid phase in situ thermal reduction method. PC-3 cells were treated with 31.25, 62.5, 125, 250, 500, 1 000 μg/ml b-TP-700 for 12, 24, 48 h. The cell activity was detected by using CCK-8 method, and the morphology of PC-3 cells was observed by using confocal fluorescence microscope, the toxicity of b-TP-700 to PC-3 cells was also judged. PC-3 cells were treated with different concentrations of b-TP-700 (31.25, 62.5, 125, 250, 500 μg/ml) and irradiated with laser (1 W/cm, 808 nm) for 1, 3, 5, 7 min. The cell activity was detected by using CCK-8 method. The production of reactive oxygen species in PC-3 cells cultured in 250 μg/ml b-TP-700 after laser irradiation for 5 min was detected by using confocal fluorescence microscopy. The non-laser irradiation cells were selected as the control group.Results:There were no statistically significant differences in cell activity of PC-3 cells treated with different concentrations of b-TP-700 for the same time ( F value was 1.26, 0.39 and 0.37, respectively; all P > 0.05). PC-3 cells were treated with 1 000 μg/ml b-TP-700 for 24 h, no obvious morphological changes and no obvious dead cells were observed under fluorescence microscope. When PC-3 cells were treated with the same concentration of b-TP-700, the activity of PC-3 cells was decreased with the extension of laser excitation time. Under the same laser excitation time, the activity of PC-3 cells was decreased with the increase of b-TP-700 concentration (all P < 0.05). Under laser irradiation, obvious green fluorescence could be detected in the PC-3 cells treated with 250 μg/ml b-TP-700, while almost no green fluorescence could be observed in the control group. Conclusions:A new photosensitizer b-TP-700 has the good reactive oxygen generation ability. b-TP-700-mediated photodynamics show obvious killing effects on prostate cancer PC-3 cells in vitro, which is expected to be a new method for the treatment of prostate cancer.
RESUMEN
Objective: To clone mouse rod opsin promoter (ROP) and establish transgenic mice harboring mouse rod opsin promoter and enhanced green fluorescent protein(mROP-EGFP) fusion gene. Methods: Mouse ROP was cloned from C57BL/6 mouse genomic DNA by polymerase chain reaction (PCR). Expression vector of mROP-EGFP fusion gene were constructed by recombination DNA technique. It was identified by restriction endonucleases digestion and confirmed by DNA sequencing. After Not I restriction endonuclease digestion, the coding elements were microinjected into male pronuclei of mice zygotes to generate transgenic mice. The pups were evaluated by PCR at genomic DNA level and mated with normal mouse. Expression of GFP in retina of transgenic mice was detected by fluorescent microscope. Results: 2. 1 kb mouse rod opsin promoter fragment was amplified from mice genome DNA. Expression vector pmROP-EGFP was constructed successfully. Following microinjection of coding sequence of pmROP-EGFP, 3 pups were verified to integrate the mROP-EGFP fusion gene in their genomic DNA by PCR assay, named C57-TgN (mROP-EGFP )SMMU21, C57-TgN (mROP-EGFP)SM-MU26 and C57-TgN(mROP-EGFP) SMMU27. They could express GFP in retina. Conclusion: 2. 1 kb mouse rod opsin promoter is cloned and expression vector pmROP-EGFP is constructed. mROP-EGFP fusion gene transgenic mice are established, which harboring mROP-EGFP gene and expressing GFP in their retina. This is valuable for studying the development of brain and retina, pathogenesis of retina disorder and retina transplanting.
RESUMEN
Objective:To evaluate the biological characters of C57-TgN(HBV adr2.0)SMMU transgenic mice. Methods: Integration,expression,replication and histology change of hepatitis B virus gene in F6 transgenic mice were estimated by ge-nomic DNA PCR,Western blotting,ELISA,immunohistochemistry,serum DNA PCR,transmission electron microscopy and H-E staining. Results: Hepatitis B virus gene was integrated into F6 C57-TgN(HBV adr2. 0)SMMU transgenic mice and expressed HBsAg,HBcAg and X protein in liver tissue. HBsAg and HBeAg were expressed in serum of 19. 54% and 3. 39% F6 transgenic mice. Hepatitis B virus were replicated in serum and liver tissue of transgenic mice. Long-term integration,expression and replication of hepatitis B virus gene induced pathological lesion of transgenic mice liver and lung. Conclusion: C57-TgNCHBV adr2. 0)SMMU transgenic mice line has the biological characters including integration of hepatitis B virus gene into genomic DNA,expression and replication of hepatitis B virus gene in serum and liver, and histological change in liver and lung. It is a valuable animal system to study pathogenesis, treatment and prevention of hepatitis B virus.