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@#This study aimed to construct a DC-targeted aptamer-modified Pseudomonas aeruginosa(PA)DNA vaccine delivery system. The cationic liposome was prepared by ethanol injection method. The cationic liposome loading pVAX1-OprF-VP22(Lip-pOprF-VP22)was prepared by electrostatic adsorption method. The encapsulation efficiency of Lip-pOprF-VP22 with different mass ratios of DOTAP/pDNA on pVAX1-OprF-VP22, cytotoxicity and transfection rate to DC2. 4 in vitro were discussed. The particle size and zeta potential of Lip-pOprF-VP22 with best mass ratio were tested. Aptamer-modified cationic liposome loading pVAX1-OprF-VP22(Apt-Lip-pOprF-VP22)was prepared by post-insertion method. The expression of OprF protein after transfection of DC2. 4 and its effect on the maturation of bone marrow-derived dendritic cells(BMDCs)were detected. Data showed that as the mass ratio of DOTAP/pDNA increased, the encapsulation efficiency of Lip-pOprF-VP22 on pVAX1-OprF-VP22 was gradually increased. When the mass ratio was 5 ∶1, pVAX1-OprF-VP22 was encapsulated well. When Lip-pOprF-VP22 with different mass ratios was applied to DC2. 4 for 24 h or 48 h, the survival rates of DC2. 4 were all above 80%. When the mass ratio of DOTAP/pDNA increased from 2 ∶1 to 10 ∶1, the transfection rate increased first and then decreased. When the mass ratios of DOTAP/pDNA were 4 ∶1 and 5 ∶1, the transfection rates were relatively high. When the mass ratio of DOTAP/pDNA was 5 ∶1, the particle size of Lip-pOprF-VP22 was(171. 67±1. 27)nm, and the Zeta potential was(11. 30±0. 57)mV. Furthermore, Apt-Lip-pOprF-VP22 can express more OprF protein and significantly promote the maturation of BMDCs. In conclusion, Apt-Lip-pOprF-VP22 can target to DC and promote the maturation of DC.
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OBJECTIVE:To investigate the early changes of ECG,cTnⅠ,LDH,α-HBDH and CKMB in patients with anthra-cyclines chemotherapy-induced heart damage after breast cancer surgery,and to explore their significances in the diagnosis of early heart damage. METHODS:Medical information of 152 cases of anthracyclines chemotherapy after breast cancer surgery in our hos-pital during Jan. 2015-Jun. 2016 were analyzed retrospectively. According to diagnosis criteria of drug-induced cardiotoxicity,the occurrence of heart damage was evaluated during hospitalization and 1-year follow-up. According to evaluation results,those pa-tients were divided into heart damage group(50 cases)and control group(102 cases). The early changes of ECG,cTnⅠ,LDH, CKMB and α-HBDH were analyzed before chemotherapy(T0),24 h after first chemotherapy(T1),24 h after second chemothera-py(T2),24 h after forth chemotherapy. RESULTS:At T1,T2,T3,the proprotion of ECG abnormalities in heart damage group was significantly higher than control group;the serum levels of cTnⅠ,LDH,CKMB andα-HBDH in heart damage group were signifi-cantly higher than control group, with statistical significance (P<0.01). CONCLUSIONS:For anthracyclines chemotherapy drugs-induced heart damage after breast cancer surgery,early regular monitoring of ECG,cTnⅠ,LDH,CKMB and α-HBDH can improve the efficiency of early diagnosis of heart damage,and improve prognosis.
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Objective To investigate the association between SLCO1B1/ApoE gene polymorphisms and lipid-lowering efficacy and safety of rosuvastatin.Methods DNA samples were extracted from blood using nano paramagnetic particle method.The SLCO1B1 521T＞C and ApoE gene polymorphisms were screened by PCR-pyrophosphate sequencing method.Totally 152 patients received rosuvastatin orally at a dose of 10 mg/d.The lipidlowering efficacy was evaluated through detecting serum low-density lipoprotein cholesterol (LDL-C) level before and 8 weeks after the treatment.The incidence of myopathic adverse effect was assessed by follow-up of the occurrence of myalgia.Results The gene distribution of SLCO1B1 521T＞C was 73.7％,23.7％ and 2.6％ respectively for TT,TC and CC in 152 patients,and the distribution of ApoE gene was 65.8％,13.2％ and 21.0％ respectively for ε3/ε3,ε3/ε2 and ε4/ε3.The genotype ε4/ε4,ε2/ε2 and ε4/ε2 were not detected.After orally receiving rosuvastatin 10 mg daily for 8 weeks,the decreased LDL-C levels showed significant differences (P＜0.05) among ApoE genotype ε3/ε2,ε3/ε3 and ε4/ε3 groups,and the frequencies of myalgia showed significant differences in the three genotype groups of SLCO1B1 521T＞C (P＜0.05).Conclusion The gene polymorphism of SLCO1B1/ApoE was correlated with efficacy and safety of rosuvastatin.The combined detection of SLCO1B1/ApoE genes can be utilized to predict efficacy and risk,and then realize individualized medication.
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Aim To compare the cytotoxicity of geni-poside (GS)and its metabolite genipin (GP)on hu-man hepatocelluar HepG2 cells and explore the sub-stance and mechanism of hepatotoxicity induced by Fructus Gardeniae.Methods The cytotoxic effect of GS and GP was analyzed by MTT method;the antioxi-dant enzyme activities of manganese superoxide dis-mutase (Mn-SOD),catalase (CAT)and levels of glu-tathione (GSH)were detected by respective kits;the change of intracellular reactive oxygen species (ROS ) was measured by 2′,7′-dichlorofluorescin diacetate (DCFH-DA)staining method;multiparameter cytotox-icity analysis (cell loss,nuclear size and morphologi-cal changes,DNA content,cell membrane permeabili-ty,mitochondrial membrane potential changes,cyto-chrome C release ) were measured simultaneously by high content screening (HCS)assays.Results No cytotoxicity was found in GS (20~1 000 μmol·L-1 ) groups (P>0.05 ),but GP was found to exert obvi-ous cytotoxic effect,and 50μmol·L-1 GP could obvi-ously inhibit HepG2 cell proliferation (P0. 0 5 ),GP could significantly decrease the activity of Mn-SOD,CAT and the level of GSH,and obviously increase the content of ROS (P0.05 ). Conclusion GP is the direct substance of hepatotox-icity induced by Fructus Gardeniae,and the mecha-nism might be associated with oxidative stress,mito-chondria injury and apoptosis.
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Objective To clarify the effects of Xinfukang containing-serum on stromal cell-derived factor-1α (SDF-1α) translation and protein secretion of bone marrow stem cells (BMSCs).Methods BMSCs were isolated and amplified using bone marrow culture method,and were identified by flow cytometry.mRNA and protein secretion of SDF-1α were detected by quantitative PCR (q-PCR) and enzyme linked immunosorbent assay (ELISA),respectively.Results The expression of SDF-1α mRNA were significantly increased after 72 h in drug-containing serum,and SDF-1α mRNA in the experimental group was approximately 200 times as that in the control group (P＜0.05).Secretion of SDF-1 α in the experimental group (277.561 1 ± 15.651 8) pg/ml was nearly doubled compared with that in the control group (153.107 1±14.765 1) pg/ml (P＜0.05).Conclusions BMSCs from whole bone marrow adherent culture have high purity,and drug-containing serum can promote BMSCs to express SDF-1 α mRNA and secretion of SDF-1 α.