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@#Objective To study the effects of attenuated Salmonella(Ty21a-pIRES-IL-2-NK4,TPIN)carrying interleukin-2(IL-2)/4-kringle antagonist of hepatocyte growth factor(NK4)double gene on humoral and cellular immune function.Methods Eighteen BALB/c mice,half male and half female,were randomly divided into control group(1. 5 mL 10%NaHCO3 by gastric tube feeding),Ty21a group(0. 1 mL Ty21a by gastric tube feeding)and TPIN group(0. 1 mL TPIN by gastric tube feeding),with 6 mice in each group. The immunization was boosted twice 7 d after the initial immunization. At 21d after administration,the blood samples were collected from eyeballs and the serum was separated,which was detected for the serum IgG antibody level by ELISA. The thymus and spleen of mice were isolated aseptically,and the spleen cells were stimulated by Ty21a and TPIN respectively in vitro. After 72 h,the proliferation ability of spleen cells was measured by CCK-8 assay,and the expression level of cytokines in spleen cells was detected by ELISA. The spleen and thymus were weighed,the spleen and thymus indexes were analyzed,and HE staining was performed.Results Compared with the control group and Ty21a group,the serum IgG level(F = 111. 74,P < 0. 01)and the contents of IFNγ,IL-4 and IL-10 in spleen cell supernatant(F = 38. 21,11. 37 and 26. 92,respectively,each P < 0. 05)increased significantly,as well as the spleen and thymus indexes(F = 10. 419 and 5. 859,respectively,each P < 0. 05)showed significant increase. In mice of Ty21a and TPIN group,the thymus cortex widened,lymphocytes increased,and there was mild inflammatory reaction;the white pulp and lymphocytes in spleen increased with neutrophil infiltration.Conclusion TPIN has a good immune protective effect,and can significantly stimulate the body to produce humoral immunity and cellular immunity,which may have a good therapeutic effect on tumors.
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@#[Abstract] Objective: To investigate the effect of 18H12, a functional monoclonal antibody that can target gastric cancer stem cells, on the self-renewal and invasion ability of gastric cancer cells. Methods: The gastric cancer cell line PAMC-82 was used as cell model, the expression of ENO1 (enolase-1) on the membrane surface of its parental cells and enriched stem cells by sphere culture was detected by Flow cytometry. Flow cytometry was used to separate ENO1+ cells and ENO1- cells to detect their self-renewal ability and invasion ability. With the commercial ENO1 antigen and antibody as the samples, CoIP (co-immunoprecipitation) was used to verify whether 18H12 antibody targeting ENO1 could able to accurately recognize ENO1. After being treated with 18H12 for 12 h, 24 h and 48 h, the selfrenewal and invasion ability of PAMC-82 cells were detected by methylcellulose pelletization experiment and Transwell chamber assay, respectively. Results: Flow cytometry showed that the expression of ENO1 on the membrane surface of PAMC-82 sphere cells was significantly higher than that of its parental cells (P<0.01), so ENO1 could be a potential target for targeting gastric cancer stem cells. The self-renewal ability and invasion ability of the sorted ENO1+ cells were significantly stronger than those of the ENO1- cells and the parental cells (P<0.05 or P<0.01). 18H12 antibody could accurately recognize ENO1, which was consistent with the commercial antibody recognition band. 18H12 could significantly inhibit self-renewal ability and invasion ability of PAMC-82 cells (P<0.01). Conclusion: Monoclonal antibody 18H12 can significantly inhibit the self-renewal and invasion of gastric cancer stem cells and is expected to be a candidate antibody drug targeting gastric cancer stem cells.
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A simple modification converts an electrospray ion source to an ambient-pressure helium plasma ionization source without the need of additional expensive hardware. Peaks for active ingredients were observed in the spectra recorded from intact pharmaceutical tablets placed in this source. A flow of heated nitrogen was used to thermally desorb analytes to gas phase. The desorption temperatures were sometimes as low as 50 1C. For example, negative-ion spectra recorded from an aspirin tablet showed peaks at m/z 137 (salicylate anion) and 179 (acetylsalicylate anion) which were absent in the background spectra. The overall ion intensity increased as the desorption gas temperature was elevated. Within the same acquisition experiment, both positive- and negative-ion signals for acetaminophen were recorded from volatiles emanating from Tylenol tablets by switching the polarity of the capillary back and forth. Moreover, different preparations of acetaminophen tablets could be distinguished by their ion-intensity thermograms.