Objective To prepare transferrin and Arg-Gly-Asp polypeptide co-modified nanoparticles(TF/RGD-NPs)and evaluate its targeting efficiency to melanoma.Methods The co-modified nanoparticles were prepared by emulsion method and its appearance,particle size and Zeta potential were evaluated.The cellular uptake experiment and melanoma tumor spheroids penetration test were used to evaluate the affinity and ability to penetrate tumor tissues of TF/RGD-NPs to melanoma B16 cells. Results The particle diameter of co-modified nanoparticles was(113.4 ±12.5)nm and the Zeta potential was(4.53 ±2.15)mV.In vitro uptake test demonstrated that the efficacy of cellular uptaken TF/RGD-NPs by B16 cells were 2.7 times and 2.9 times to TF-NPs and RGD-NPs,respectively,the differences were all significant(P<0.05 ).Tumor spheroid penetration test results showed that TF/RGD-NPs has good affinity to melanoma cells.Conclusion TF/RGD-NPs can target to melanoma B16 cell efficiency in vitro,it may be serve as a potential drug delivery system for targeting melanoma.

Objective To explore the effect of X-linked inhibitor of apoptosis protein(XIAP)combined with doxorubicin on the proliferation of MCF-7 cell and growth of MCF-7 bearing nude mice.Methods The anti-proliferation efficiency of doxorubicin and embelin were determined by MTT assay.MCF-7 cell apoptosis induced by embelin and doxorubicin were detected by Flow cytometry.Anti-tumor ability of embelin and doxorubicin were evalued with tumor spheroids test.MCF-7 cell were xenografted to mice to establish the animal model,which was used to evaluate the effect of anti-tumor. Results Compared with saline group,embelin group and doxorubicin group,the combination(doxorubicin +embelin)group inhibited the growth of MCF-7 cells effectively(P<0.01).The combination group induced the apoptosis of MCF-7 cells more effectively than doxorubicin alone(P<0.01), and significantly inhibit the growth of tumor in vitro and in vivo than other groups(P<0.01).Conclusion The combination of doxorubicin and embelin may be used as a potentially effective treatment method for breast cancer.

Acute Disease ; Animals ; Charcoal ; therapeutic use ; Enema ; Milk ; Paraquat ; toxicity

Objective In this article we evaluated the sensitivities and specificities of real-time PCR assay for diagnosis of human brucellosis.Methods The species selectivity and specificity of real-time PCR were evaluated by direct amplification of a 169 bp portion of bcsp31 gene from 15 Brucella strains and 41 non-Brucella strains.According to the monitoring results of 2012 Harbin brucellosis,17 brucellosis patients and 30 health people were selected to collect their serum samples for assessing the sensitivity of real-time PCR,and additional 30 nonbrucellosis patients serum samples were as controls.Results The species selectivity and specificity of our realtime PCR method were evaluated by using genomic DNA from 15 Brucella strains and 41 non-Brucella strains.There were 11 sera with positive amplification signals among the 17 culture-proven brucellosis patients,the sensitivity was 64.7％(11/17).Whereas,the results of sera from the 60 control patients were all negative,corresponding to a specificity of 100.0％.Conclusion The results indicate that real-time PCR is well suitable for confirmation of brucellosis cases.

Objective As a signaling molecule,NO regulates key physiological processes and is closely related to periodontitis.To investigate the effect of flavonoid NO donor composite nanoparticles(G10@HAP/MSN@ZnO@COS)on osteogenic differentiation of periodontal ligament stem cells(PDLSCs)by regulating macrophage polarization.Methods The novel NO donor drug G10 was loaded on hydroxyapatite/mesoporous silicanant particles(HAP/MSN),filled with zinc oxide(ZnO),and then coated with chitosan(COS)to prepare composite nanoparticles(G10@HAP/MSN@ZnO@COS).The best concentration of G10@HAP/MSN@ZnO@COS was screened to promote cell proliferation by CCK-8 cell experiment.After the mouse mononuclear macrophages were stimulated by lipopo-lysaccharide,the mice were divided into four groups:Control group,G10 group,HAP/MSN@ZnO@COS group and G10@HAP/MSN@ZnO@COS group.Each group was cultured with fresh medium,5 μg/mL G10,5 μg/mL HAP/MSN@ZnO@COS and 5 μg/mL G10@HAP/MSN@ZnO@COS for 72 h respectively.ELISA and RT-qPCR were used to detect the expression of cytokines(TNF-α,IL-6,IL-1β,iNOS,IL-10)and mRNA expression in each group,and the phenotypic changes of M1/M2 were evaluated.The supernatant of each culture medium was used as conditioned medium to culture PDLSCs,and the osteogenic ability and cell miner-alization were evaluated by alkaline phosphatase activity test and alizarin red staining.Results CCK-8 experiment showed that G10@HAP/MSN@ZnO@COS of 5 μg/mL could significantly promote the proliferation of PDLSCs.The results of ELISA showed that compared with Control group,the expression of M1 type marker IL-1β,IL-6,TNF-α and iNOS in G10@HAP/MSN@ZnO@COS group was significantly decreased(P＜0.000 1),while the expression of M2 type marker IL-10 was significantly increased(P＜0.000 1).The results of RT-qPCR were consistent with those of ELISA,which showed that the expression of M1-related genes in G10@HAP/MSN@ZnO@COS group decreased significantly(P＜0.01).The results of alizarin red staining and alkaline phosphatase activity test showed that the number of mineralized nodules and alkaline phosphatase activity in G10@HAP/MSN@ZnO@COS-CM group were significantly higher than those in other groups(P＜0.000 1).Conclusion Composite nanoparticles(G10@HAP/MSN@ZnO@COS)can effectively inhibit the polarization of macrophages to M1 phenotype and promote it to M2 phenotypic polarization.The anti-inflammatory microenvironment regulated by G10@HAP/MSN@ZnO@COS can en-hance the osteogenic differentiation of PDLSCs.

Objective: To investigate the effect of Wnt3a on osteogenic differentiation of human dental pulp stem cells (DPSC). Methods: DPSCs were subjected to different concentrations of Wnt3a (0, 5, 20, 50 and 100 μg/L) and at seven days after culture the alkaline phosphatase (ALP) activity was tested. Mineralized nodule formation was examined by alizarin red staining. Osteogenic-related gene expression of bone sialoprotein (BSP), osteocalcin (OCN), collagen type Ⅰ (COL-Ⅰ), Runt-related transcription factor-2 (RUNX2) was examined by quantitative real-time PCR (qPCR). Results: After seven days of induction by DPSC, Wnt3a protein could inhibit the ALP activity (concentration 0: 1.076±0.203, 5 μg/L: 0.828±0.118, 20 μg/L: 0.505±0.044, 50 μg/L: 0.499±0.038, 100 μg/L: 0.483±0.060). The expression of OCN in 5 μg/L Wnt3a group (0.092±0.005) was lower than that in culture medium (0.858±0.190)(P<0.05). Alizarin red staining showed that 5 μg/L Wnt3a had no mineralization induction effect on DPSC. Conclusions Wnt3a could inhibit osteogenic differentiation of dental pulp stem cells.