1.Synthesis of silver carboxymethyl chitosan and its experimental study on its bacteriostasis
Xuejun ZHAN ; Yuanzhen XIONG ; Zhe LIU ; Daze XIE
Chinese Journal of Biochemical Pharmaceutics 2001;22(3):142-144
Purpose The aim is to develop a synthesis method of the silver carboxymethyl chitosan and to study its bacteriostasis to Staphlococcus aureus(S.aureus),Pseudomonas aeruginusa(P.aeruginusa),Escherichia Coli(E.coli),Klebsiella pneumoniae(K.pneumoniae) and Proteus vulgaris(P.vulgaris).Methods Chitosan was modified by way of chemistry.The structure analysis of its derivate was analysed by infra-red absorption spectroscopy,using methods of dilution and concavering to study the bacteriostasis to some ordinary bacteria which cause infection in burn.Results The infra-red spectragram of the derivate showed the chitosan had been modified by chloroacetic acid.When the concentrationl of silver carboxymethyl chitosan was 1.028 mg/ml and the concentration of the five bacteria were 104CFU/ml,the rate of bacteriostasis was 88%、80.2%、75.3% to S.aureus、P.aeruginusa and E.coli respectively.The MIC of silver carboxymethyl chitosan was similar to that of AgNO3 when they act on S.aureus and P.aeruginusa,but the former was lower than later when acting on E.coli.Conclusion Silver carboxymethyl chitosan could inhibit some bacteria which caused infection in burn.It was a novel pharmaceutical in preventing and curing burn infection.
2.Effect of LX A4 and BML-111 on TLR4/NF-κB signaling pathway in macrophage
Daze XIE ; Lixing HUANG ; Dongsheng LIU ; Jun ZHU ; Yong XIE ; Nanjin ZHOU
The Journal of Practical Medicine 2015;(17):2799-2802
Objective To determine the effect of lipoxins (LX) A4 and its agonist (BML-111) on the survival of RAW264.7 macrophage cells and TLR4/NF-κB signaling pathway. Methods RAW264.7 cells were treated with different concentrations of LPS, then the effect of LX A4 and BML-111 on the survival rate of these cells was observed. Cytotoxicity were detected by CCK-8 method and RT-PCR was used to detect the TLR4 and TRAF6 mRNA. The protein levels of TLR4 and pNF-κB p65 in RAW264.7 were determined by Western Blot. Results The survival rates of macrophage treated with LPS for 6 h in 1 000 ng/mL LPS in LX A4 group and BML-111 group were significantly higher than that in control group (P < 0.05). In the present of LPS, the TLR4 mRNA levels in RAW264.7 cells from LX A4 group and BML-111 group were significantly higher than those in the corresponding non-LPS groups. And the TRAF6 mRNA levels in each LPS stimulation group were higher than those in the corresponding non-LPS groups (P<0.05), while the protein level of TRAF6 in LX A4 and BML-111 groups were significantly lower than that in control group (P < 0.05). Stimulated with LPS, the protein levels of pNF-κB p65 in the LX A4 group and BML-111 group were all significantly lower than that in control group (P<0.05), and pNF-κB p65 expression level in control group was also significantly higher than the corresponding non-LPS groups (P < 0.05). Meanwhile, no significant difference was found between LX A4 and BML-111 group (P > 0.05). Conclusion LX A4 and BML-111 could inhibit the cytotoxicity of LPS on the RAW264.7 macrophage cells through the inhibition of the activation of TLR4/NF-κB signaling pathway, then reduce the inflammation. And this stable BML-111 may appear as another promising treatment for IBD disease.
3.Effect of triptolide on human colorectal cancer HCT116 cell proliferation, autophagy and apoptosis
Lin ZHAO ; Peng WU ; Pinggui ZHANG ; Daze XIE ; Dian GAO ; Nanjin ZHOU
Chinese Pharmacological Bulletin 2016;32(10):1399-1403,1404
Aim To investigate the effect of triptolide ( TP) on human colorectal cancer HCT116 cell prolif-eration and explore the potential mechanism of TP in treating colon cancer. Methods MTT assay was used for estimating the survival rates of HCT116 cells ex-posed to different concentrations and different duration time of TP. Western blot ( WB ) was used for testing the expression of LC3-Ⅱ. MDC staining was employed to detect cell autophagy. Flow cytometry ( FCM) was applied to test the effects of TP on cell apoptosis. Re-sults TP could inhibit cell proliferation in HCT116 cells. The expression of LC3-Ⅱwas enhanced with the increase of the concentration of TP after exposed to dif-ferent concentrations of TP for 48 h and the duration time of TP after exposed to 40 nmol·L-1 TP,and the number of acid autophagic vacuoles in HCT116 cells had increased, which was observed by fluorescence mi-croscope. The HCT116 cells apoptotic rates in TP group had decreased compared to control group and de-creased significantly compared to TP +3-MA group. The HCT116 cells apoptotic rates in TP group had in-creased compared to control group and increased signif-icantly compared to TP+RAPA group. Conclusion TP could inhibit the proliferation of human colon canc-er HCT116 cells and induce apoptosis in HCT116 cells, which indicates that synergy may exist between autophagy and apoptosis.
5.Preparation and immunological evaluation of oral solution of egg yolk-derived hepatitis B virus-specific transfer factor.
Yanping XU ; Qing ZHANG ; Xuejun ZHAN ; Daze XIE ; Ge DAI ; Hui YANG
Journal of Southern Medical University 2013;33(12):1827-1830
OBJECTIVETo prepare the oral solution of egg yolk hepatitis B virus (HBV)-specific transfer factor (EYHBV-TF) and evaluate its immunological activity as an immune regulator against hepatitis B.
METHODSFrom hens immunized with the Hepatitis B vaccine the egg yolk was isolated to extract the specific transfer factor EYHBV-TF, and its physicochemical properties were examined. Leukocyte adhesion inhibition test (LAI) was performed to detect the immunogenic activity of EYHBV-TF. The solution of EYHBV-TF was then administered orally in normal mice, and the specific cellular immune activity induced was assayed with delayed type skin hypersensitivity test (DTH), with the non-specific immune activity assessed with immune organ index. The immune responses induced by oral EYHBV-STF solution were compared with those by EYHBV-STF injection and by different dosages (injection and oral) of porcine spleen HBV-specific transfer factor (PSHBV-STF), porcine spleen nonspecific transfer factor, and egg yolk extracts from non-immunized hens.
RESULTSThe prepared EYHBV-STF oral solution, which met the standards for biological products, could inhibit leukocyte adhesion in vitro and significantly enhance mouse foot pad swelling, demonstrating its capability of transferring antigen-specific delayed type hypersensitivity reactions to naive recipient. EYHBV-STF oral solution also significantly improved the immune organ index in mice (P<0 01) with similar effects to those caused by EYHBV-STF injections and by PSHBV-STF injection and oral solution.
CONCLUSIONOrally administered EYHBV-STF and EYHBV-STF injection both possess hepatitis B antigen-specific cellular immune activity and can significantly enhance specific cellular immune responses.
Animals ; Chickens ; Egg Yolk ; chemistry ; Hepatitis B ; drug therapy ; Hepatitis B Antigens ; Hepatitis B virus ; drug effects ; Immunity, Cellular ; Immunization ; Mice ; Swine ; Transfer Factor ; administration & dosage ; pharmacology