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Objective To analyze the physicochemical properties and photothermal properties after introducing a certain proportion of phaeophorbide into polyvinyl alcohol-lipoic acid polymer.Methods The "one-step" method was adopted to synthesize the PVA27K-4 ％LA-10 ％ Pheide (PLP) polymer.The laser particle diameter tester was used to detect the particle size,polydispersity index and ζ potential of PLP nanoparticle (PLPN);the morphology was observed under transmission electron microscope (TEM);the in vitro fluorescent and photothermal properties were investigated by using spectrophotometric method and infrared thermal imager;the ovarian-cancer nude mice models were established and the preliminary in vivo experiment evaluation was performed.Results The synthetic PLP polymer could self-assembly form spherical PLPN in aqueous environment,which showed the tightly spherical structure under TEM.The average particle diameter,polydispersity index and ζ potential were (22.92 ± 9.00)nm,(0.117±0.056) and (-25.43±0.77)mV respectively.The fluorescence intensity of PLPN in PBS solution was significantly lower than that in SDS solution (P＜0.05).However,the temperature of the PBS solution of PLPN was significantly higher than that of the SDS solution after exposure to the near infrared light (P＜0.05).In the model of tumor-bearing nude mice,PLPN could targetaccumulated in the tumor site.The temperature in the tumor site was significantly increased after exposure to the near infrared light,which could kill tumor cells.Conclusion PLPN could emit red fluorescence,and could target-accumulated in tumor site and conduct the in vivo image,which has significant photothermal conversion properties in vivo and in vitro,and can effectively inhibit the tumor growth in vivo.
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BACKGROUND:Caffeic acid phenethyl ester (CAPE) can inhibit lipid peroxidation after rat brain injury. However, the trend of 5-lipoxygenaseis (5-LOX) and cysteinyl leukotrienes (CysLTs) in model of Parkinson’s disease, and whether CAPE protects against rotenone-induced cel ular injuries by inhibiting the levels of 5-LOX and CysLTs stil need further research. OBJECTIVE:To investigate the protective effect of CAPE on the rotenone-induced Parkinson-like injury, and to determine whether 5-LOX involved. METHODS:(1) PC12 cel s in good-growth were col ected and divided into five groups cultured with different concentrations of rotenone (0, 0.01, 0.1, 1, 10μmol/L). 24 and 48 hours later, changes of cel ular morphology and activity were observed to single out the optimum concentration of rotenone;at 24 hours, the levels of 5-LOX and CysLTs were detected by western blotting and ELISA, respectively. (2) PC12 cel s were pretreated with different concentrations of CAPE (0, 0.01, 0.1, 1, 10 μmol/L) for 30 minutes, and 1 μmol/L rotenone was then added. The other cel s received no intervention as blank control group. Subsequently, the cel activity was detected, and the CysLTs production was detected by ELISA at 24 hours. RESULTS AND CONCLUSION:(1) Rotenone (0.1-10μmol/L) could induce PC12 cel injury with overt morphological and cel activity changes at 24 hours, especial y the 1 μmol/L rotenone. (2) Rotenone also significantly increased the 5-LOX expression and CysLTs production in a concentration-dependant manner. (3) CAPE (1-10μmo/L) significantly attenuated rotenone-induced CysLTs production and cel viability reduction in a concentration-dependant manner. (4) These results suggest that CAPE protects against PC12 cel injuries in the model rat with Parkinson’s disease induced by rotenone involving 5-Lox.
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<p><b>BACKGROUND</b>p16INK4a and p14ARF, encoded by gene INK4a/ARF located at chromosome 9p21, are cyclin dependent kinase (CDK) inhibitors. Both p16INK4a and p14ARF are cell cycle regulatory proteins and play an important role in Rb and p53 passways respectively. In this study, wild-type INK4a/ARF gene was transfected into human lung adenocarcinoma cell line A549, in which this gene site was lost, and the effects on the cell's biological behavior were investigated.</p><p><b>METHODS</b>The recombinant eukaryotic expression plasmids pcDNA3-p16INK4a and pcDNA3-p14ARF were transfected into A549 by cationic liposome method. By RT-PCR, immunocytochemistry and Western blot after G418 selection, A549 cells that could stably express p16INK4a and p14ARF were obtained. As a control, the parental cell and negative control cell with plasmid pcDNA3-LacZ were used. Inhibition of proliferation was measured by MTT assay. The cell growth curve was drawn according to cell counts. Cell cycle distribution was measured by flow cytometry (FCM), the apoptosis indexes were observed at the same time. The colony formation rate was counted by staining the cells with Coomassie brilliant blue.</p><p><b>RESULTS</b>The introduction of exogenous INK4a and ARF caused significantly growth inhibition of A549. By FCM, more percentage of A549-p16INK4a-p14ARF cells couldn't pass through the checkpoint G1. The percentage of A549-p16INK4a-p14ARF cells inhibited at G0/G1 was 59.9%, 50.3% for A549-vector and 51.2% for A549. The statistical differences were significant between A549-p16INK4a-p14ARF cell and A549-vector cell (P=0.025) and between A549-p16INK4a-p14ARF cell and A549 cell (P=0.043). The apoptosis index of A549-p16INK4a-p14ARF cell was 8.0% and 2.7% for both A549-vector and A549 cell (P < 0.01). The colony formation ability of A549-p16INK4a-p14ARF was weaker than that of A549-vector and A549, they were 63%, 87% and 85% respectively.</p><p><b>CONCLUSIONS</b>The wild-type INK4a/ARF gene can be co-introduced effectively into A549 cell by cationic liposome method. The reexpression of p16INK4a and p14ARF in A549 can inhibit the growth and enhance the apoptosis. This trial will be helpful in using gene therapy of lung cancer in the future.</p>
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Objective To investigate the influence of co-expression state of p14 ARF and p16 INK4a protein on radiochemotherapy and length of survival period in patients with non-small cell lung cancer(NSCLC)after resection. Methods Thirty-five patients with negative p14 ARF and p16 INK4a co-expression and 20 patients with positive p14 ARF and p16 INK4a co-expression were enrolled for the study. The co-expression of the said proteins were previously determined by immunohistochemistry (S-P). Clinical pathological characteristics were compared between two groups, and the survival time and the results of radiochemotherapy of patients were respectively recorded and analysed. Results No significant differences were found in age, TNM stages, degree of differentiation, recurrence/metastasis and radiochemotherapy between two groups. However, there was significant differences in sex, smoking index, and pathological classification. It was found that 2-year, 3-year and total survival rate were significantly lower in patients with p14 ARF and p16 INK4a co-expression than those with positive co-expression (P