Polypeptide-rhodamine B probes containing laminin/fibronectin receptor-targeting sequence (YIGSR/RGD) for fluorescent imaging in cancers.

Currently, fluorescent imaging is one of the most promising diagnostic approaches for facile detection of cancers in situ in thanks to a fluorescent probe. Two novel polypeptide-based fluorescent probes for different biomarkers to cancers are reported here. These probes focused on tyrosine-isoleucine-glycine-serine-arginine (YIGSR) and arginine-glycine-aspartic (RGD), which receptors play an important role in the extracellular matrix and are overexpressed in tumor cells and then can be used as tumor-targeting groups in fluorescent imaging. In this work, the pentpeptide-rhodamine B derivative (YIGSR-RhB) and tripeptide-rhodamine B derivative (RGD-RhB) were synthesized respectively by using the solid phase synthesis methods. These derivatives were further characterized by 1HNMR, MS, UV and IR, etc. Their fluorescent and biocompatibility properties, such as the cell cytotoxicity, cell uptake and fluorescent imaging of tumor cells, and fluorescent imaging in BALB/c female mice with 4T1 tumors and C57 mice with B16F10 tumor in vivo, were also measured. Experiment results demonstrated that YIGSR-RhB and RGD-RhB possessed the low cell cytotoxicity, good tumor-targeting property and fluorescent properties similar to rhodamine B. Moreover, YIGSR-RhB and RGD-RhB can be taken up highly by the B16F10 melanoma cells and 4T1 breast cancer cells, and then achieve the good fluorescent imaging in these tumor cells in vitro and tumors of mice in vivo. Therefore, YIGSR-RhB and RGD-RhB can be used as the potential tumor-targeting probes for fluorescent imaging. They can directly attach the cell membrane and specifically target to the tumor cells.

[Establishment and application of an intact natural model of human dental plaque biofilm].

PURPOSE: Human dental plaque is implicated in a number of oral diseases. Collection of its undisturbed intact structure facilitates observing the formation and the effect of treatment. This study established a model in vivo to collect intact natural dental plaque biofilm, and observed the features and evaluated the immediate penetration and bactericidal effect of an essential oil (EO) mouthrinse on it at different periods. METHODS: Three 500 microm wide grooves were cut into hydroxyapatite (HA) discs. The discs were worn by six volunteers for 6, 24 and 48 hours, then broken into halves, one served as control, while the other received a one-minute extra-oral EO mouthrinse treatment. 5, 15 and 30 minutes later, the plaque was visualized with a vitality staining technique to observe the sustained changes of biofilm structure in situ and the effect of EO on intact biofilm. The biofilm thickness and fluorescent density of vital and total bacteria were obtained through diagram analysis, assessing the percentage of thickness and fluorescent density of vital to total bacteria. All data analysis was performed using SPSS 13.0 software package. One-way ANOVA and Student-Neuman-Keuls tests were used. RESULTS: The thickness of 6h, 24h and 48 h biofilm was (11.92 + or - 4.63) microm, (18.63 + or - 4.66)microm, (27.55 + or - 6.35) microm, respectively, which increased significantly within 48 hours (P<0.05), especially those within 6 hours. The percentages of thickness and fluorescent density of vital to total bacteria at different periods showed no significant changes (P>0.05). For 6-hour samples, plaque vitality for thickness and fluorescent density decreased significantly within 5 minutes after exposure to the EO (P<0.05). Meanwhile, for 24, 48-hour samples, plaque vitality decreased significantly within 15 minutes (P<0.05). CONCLUSIONS: This study successfully establishes a model for collecting natural undisturbed plaque biofilm in situ, showing the changes constantly within 48 hours. EO mouthrinse has an immediate penetration and antimicrobial effect on the sustained dental plaque biofilm, especially 15 minutes later.

[Protective effect of tert-butylhydroquinone on bone marrow cells in rats from cytotoxicity induced by benzene in vitro].

OBJECTIVE: To investigate the protective effect of tert-butylhydroquinone on bone marrow cells in rats from cytotoxicity induced by benzene in vitro. METHODS: The bone marrow cells in rats were divided into two groups randomizedly. Cells of the control group were stimulated by 0, 5, 10, 15, 20 mmol/L benzene for 2, 4, 6 hours respectively. Cells of the tBHQ-pretreated group were treated by 100 micromol/L tBHQ for 12 hours followed by the same conditions as the control group. The DNA damage was detected by single cell gel electrophoresis assay (SCGE) and cell apoptosis was examined by flow cytometry. The activities of NAD (P) H: quinone oxidoreductase (NQO1) in bone marrow cells of rats were also measured before benzene treatment in two groups. RESULTS: In control group, the DNA damage and the apoptosis of bone marrow cells was increased with the growing concentration and time of benzene treatment. The DNA migration and the lengths of DNA migration of the bone marrow cells in the rats under 5, 10, 15, 20 mmol/L benzene treatment in the tBHQ-pretreated group were significantly lower than those in control group at the same time point (P < 0.05). The apoptosis of the bone marrow cells in the rats stimulated by 15, 20 mmol/L benzene for 2 hours and 10, 15, 20 mmol/L benzene for 4 hours as well as 5, 10, 15, 20 mmol/L benzene for 6 hours were also significantly lower than those in control group (P < 0.05). The activities of NQO1 in the bone marrow cells in the rats were increased after tBHQ treatment (P < 0.01) (1.62 +/- 0.16 min(-1).mg(-1) vs. the control group: 0.95 +/- 0.08 min(-1).mg(-1)). CONCLUSION: The benzene can induce the DNA damage and the apoptosis of bone marrow cells in rats in a time dependent and dose dependent manner to some extent. The tBHQ can protect the bone marrow cells in rats from the cytotoxicity induced by benzene, which can be partly explained by the increase of the NQO1 activity induced by tBHQ.

CdSe/ZnS-labeled carboxymethyl chitosan as a bioprobe for live cell imaging.

A simple and convenient method for the construction of CdSe/ZnS-labeled polysaccharides as bioprobes were developed, which are highly biocompatible and photostable, and have been proven to be suitable for live cell imaging.