Object To investigate the stability of baicalin in different conditions Methods The stability of baicalin in water with different pH values, methanol, ethanol, chloroform and in the mouse lung homogenate, mouse liver homogenate, mouse plasma and bovine serum was investigated by the classical isothermal method Results The experiments showed the baicalin was stable in acidic water and these organic solutions, but unstable in basic water and these biological media Conclusion The stability of baicalin in different media was different, which should be considered in practice

Objective: In order to provide an effective reference to the study on its preparation, the stability in vitro of andrographolide was investigated. Methods: Andrographolide was dissolved in phosphate buffer solutions of different pH values; classical isothermal method was employed; the content of andrographolide was determined by RP HPLC; then the stability of the drug in different biological media and organic solvents at 37?C was investigated. Results: The stability of andrographolide was different at different pH values and was best in the condition of pH=3～5. In biological media, it was most stable in bovine serum and the next was in mice homogenate. Conclusion: In different conditions, the stability of andrographolide was different. It provided a reference to the study on andrographolide and its preparation.

After percutaneous coronary intervention (PCI) was performed,since CHD risk factors still exist, coronary restenosis rate remains high.Therefore, self-management after PCI is very important.The present article made a review on current condition and research progress of self-management in patients after PCI, aiming at providing reliable evidence for rehabilitation after PCI.

PurposeTo investigate the image quality of Dual Source CT (DSCT) high pitch prospective scan in acute chest pain examination by using triple-rule-out (TRO) protocol and its responses to heart beat rate change.Materials and Methods Thirty-two consecutive patients who planned to undergo TRO acute chest pain examination were enrolled and randomly divided into research group and control group. The conventional retrospective spiral scanning protocol was applied to the patients in the control group, while the high pitch prospective spiral scanning mode was used in the research group. CT value, noise level, SNR, CNR and radiation dosage in aorta and pulmonary arteries were recorded and compared between the two groups, and the image quality of the coronary arteries was scored subjectively.Results No significant difference was found in the aspects such as CT value, noise level, SNR, CNR radiation dosage in aorta and pulmonary arteries (P>0.05); however, the subjective scoring for coronary artery in the research group (85.5%) was significantly lower than that in the control group (93.9%) (χ2=8.33,P<0.05). Considering the influence of severe heart rate changes, we excluded those patients whose heart rate variance was more than 5 beats/min (control group:n=5; research group:n=4) and then compared the image quality of the two groups again. The subsequent results showed no difference (χ2=0.12,P>0.05). In addition, the radiation dosage in the control group was significantly higher than that in the research group [(20.8±6.2) mSvvs. (2.3±0.4) mSv;t=17.142,P<0.01].Conclusion DSCT high pitch prospective scanning in TRO protocol shows high success rate in examining patients with acute chest pain whose heart beat rate is lower than 80 beats/min and heart beat variance is less than 5 beats/min. The image quality has no difference with that derived from retrospective scan but with far more less radiation dose (2.3±0.4) mSv. This method thus is promising and can be widely promoted clinically.

Lysine-specific demethylase 1 (LSD1) plays vital roles in cell stemness, differentiation, cell motility, metabolic control and epithelial-mesenchymal transition, which is closely associated with tumorigenesis processes including cell proliferation, invasive, metastasis and poor prognosis. Besides, LSD1 also contributes to the occurrence of other diseases such as neurodegenerative diseases and viral infections. Since 2013, the irreversible inhibitors including tranylcypromine, ORY-1001, ORY-2001, GSK-2879552, IMG-7289, INCB059872, TAK-418, LH-1802 and reversible inhibitors including CC-90011 and SP-2577 have been approved for clinical assessment. This review comprehensively summarizes the clinical research of LSD1 drug candidates and briefly discusses the prospects, opportunities and challenges of LSD1-targeted drug discovery, aiming to provide a landscape for the related drug development.

Boronic Acids ; therapeutic use ; Bortezomib ; Humans ; Male ; Middle Aged ; POEMS Syndrome ; therapy ; Peripheral Blood Stem Cell Transplantation ; Pyrazines ; therapeutic use ; Transplantation, Autologous

Objective To study the role of protective antigen(PA)and N-terminal segment of lethal factor (LFn)in the entrance of EGFP(enhanced green fluorescent protein)into HeLa cells. Methods The DNA fragments encoding LFn and EGFP were amplified,respectively,and cloned into the plasmid pET-21 a(+)one after another to construct a recombinant plasmid pET-LFn-EGFP. The plasmid was txansformed into BL21 cells to express LFn-EGFP protein under the induction of IPTG. The protein was purified by Ni chelating chromatography. After incubation with LFn-EGFP in the presence of PA or not, the HeLa cells were analyzed by flow cytometry or laser confocal microscopy. Results The fusion protein LFn-EGFP was purified by over 90% homogeneity and retained the ability of LF to bind with PA when incubated with J774A.1 macrophage cells,and could get into HeLa cells. Conclusion The LFn-EGFP could enter the HeLa cells in a PA independent pathway. But PA could help more LFn-EGFP molecules enter into HeLa cells.

Recombinant Fl-V (rFl-V) fusion protein is the main ingredient of the current candidate vaccine against Yersinia pestis infection, which has been under investigation in clinical trial in USA. We investigated the soluble expression conditions of rF1-V in Escherichia coli BL21 (DE3) that we constructed before. After scale-up and optimization of fermentation processes, we got the optimized fermentation process parameters: the culture was induced at the middle exponential phase with 50 µmol/L of IPTG at 25 °C for 5 h. Soluble rFl-V protein was isolated to 99% purity by ammonium sulfate precipitation, ion exchange chromatography, hydrophobic chromatography and gel filter chromatography. The protein recovery was above 20%. Protein identity and primary structure were verified by mass spectrometry and Edman sequencing. Results of purity, quality and western blotting analysis indicated that the target protein is a consistent and properly folded product. Furthermore, the immunogenicity of various antigens formulated with aluminum hydroxide adjuvant was evaluated in mice. Serum antibody titers of 4 groups including 20 µg rFl, rV and rFl-V and 10 µg rFl+10 µg rV, were assayed by ELISA after 2 doses. The antibody titers of anti-Fl with 20 µg rFl-V were obviously higher than titers with other groups; meanwhile there were no significant difference of anti-V antibody titers among them. These findings confirm that rFl-V would be the active pharmaceutical ingredient of the plague subunit vaccine.
Adjuvants, Immunologic ; Animals ; Antibodies, Bacterial ; blood ; Antibody Formation ; Antigens, Bacterial ; immunology ; Blotting, Western ; Chromatography, Ion Exchange ; Enzyme-Linked Immunosorbent Assay ; Mice ; Plague ; prevention & control ; Plague Vaccine ; immunology ; Recombinant Fusion Proteins ; immunology ; Vaccines, Subunit ; immunology ; Yersinia pestis

BACKGROUND:Reports have demonstrated that cytotoxicity produced by ferric oxide (Fe2O3) nanoparticles is associated with cellular lipid peroxidation. Whether Fe2O3 nanoparticles have toxicity to macrophages, and what is the association of toxic mechanism and oxidization?OBJECTIVE: To observe the effects of different concentrations of Fe2O3 nanoparticles on the oxidative damage of macrophages.DESIGN: A controlled observation experiment.SETTING: School of Public Health, Southeast University.MATERIALS: RAW264.7 cells were peritoneal macrophages of mouse and purchased from Shanghai Institute of cells, Chinese Academy of Sciences. Fe2O3 nanoparticles (30 nm) suspension was provided by Department of Biomedical Engineering, Southeast University). Fe2O3 nanoparticle suspension was placed in 60 ℃ water for 10 hours,then in 37 ℃ water overnight. This procedure was repeated 3 times for germicidal treatment. Then, the suspension was packed into small bottles and stored at 4 ℃ for later use. DMEM high glucose culture fluid (Gibco Company,USA); trypsinase (Difco Company, USA, imported); new-bom calf serum(Sijiqing Company, Hangzhou); hydrogen dioxide (H2O2, Gibco Company); Kits for measuring hydrogen dioxide(H2O2), hydroxy radical (·OH), superoxide anion radical (O2·-), lactic acid dehydrogenase, ultramicro ATP enzyme and Coomassie brilliant blue protein levels (Jiancheng Biotechnique Co., Ltd.,Nanjing).METHODS: This experiment was carried out in the laboratory of Department of Labor and Environmental Health, School of Public Health, Dongnan University between March 2006 and July 2006. RAW264.7 cells (Abelson murine leukemia virus-induced tumor) were cultured in DMEM (Gibco Company) containing 100 g/L fetal bovineserum, 100 000 U/L penicillin and 100 mg/L streptomycin in the environment of 5% CO2. Cell growth was observed under an inverted radical in the cells: 1.5×108 L-1 macrophages were inoculated to 24-well plate, 1 mLa well. After the macrophages were cultured for 24 hours in incubation at 37 ℃ in a humidified atmosphere containing 5% CO2. 1.070 0, 0.5350 and 0.2675 g/L Fe2O3 nanoparticles (30 nm) suspension-intervened macrophages were set as Fe2O3 nanoparticle group, and normal saline group was set as control group. Following the intervention of nanoparticles, macrophages were disrupted with Determination of the activities of lactate dehydrogenase (LDH), Na+-K+-ATPase and Ca2+-Mg2+-ATPase: Macrophages in the Fe2O3 nanoparticle group and control group were treated as above. The activities of LDH in culture medium were determined according to the instruction of reagent kit (Nanjing Jiancheng Bioengineering Co., Ltd). And the activities of Na+-K+-ATPase and Ca+-Mg2+-ATPase were also determined according to the instruction of reagent kit (Nanjing Jiancheng Bioengineering Co., Ltd) at low temperature. MAIN OUTCOME MEASURES: ①Effects of different concentrations of Fe2O3 nanoparticles on the production of H2O2, ·OH and O2·- in RAW264.7 cells.②Effects of different concentrations of Fe2O3 nanoparticles on the activities of LDH ,Na+-K+-ATPase and Ca2+-Mg2+-ATPase in RAW264.7 cell culture fluid.RESULTS: ① Level of ·OH free radical in Fe2O3 nanoparticle 0.267 5, 0.535 0, 1.070 0 g/L groups was higher than that in control group, respectively [(0.605±0.066), (0.410±0.080), (0.764±0.051), (0.285±0.057)mkat/g, P ＜ 0.05]; Level of respectively [(9.935±1.159), (8.912±0.131), (13.479±0.752), (5.635±0.475)μkat/g,P ＜ 0.05]; Level of H2O2 in Fe2O3 nanoparticle 1.070 0 g/L group was higher than that in the control group [(14.695±2.815), (2.397±0.399) mmol/L, P ＜increased (P ＜ 0.05). Fe2O3 nanoparticles had effects on the activities of Na+,K+-ATPase and Ca2+,Mg2+-ATPase. With the increase of dose of Fe2O3 nanoparticles, the activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase were gradually decreased. There were significant differences as compared with control group (P ＜ 0.05)CONCLUSION:Increasing dose of Fe2O3 nanoparticles wouldcause more H2O2,·OH and O2·- free radicals in the cells, increase cell membrane permeability and inhibit the activities of LDH, Na+-K+-ATPase and Ca2+-Mg2+-ATPase.

Objective:To observe the effect of pecking moxibustion on the pattern characteristics of redness,swelling,heat,and pain in the affected joints,also the synovial cell ultrastructure in rats with rheumatoid arthritis(RA)due to damp heat affecting bones/joints,and to explore the anti-inflammatory mechanism of pecking moxibustion in treating the early stage of RA. Methods:Eighteen rats were randomly selected from 78 female ones as the blank group,and all the other rats were subjected to preparing the"differentiation of disease and pattern"RA model due to damp heat affecting bones/joints by using the method of"collagen-induced arthritis plus windy,damp,and hot environment stimulation".Fifty-four rats with successful modeling were randomly divided into a model group,a drug group,and a pecking moxibustion group,with 18 rats in each group.Rats in the drug group were given methotrexate at a dose of 1 mg/(kg·bw)on the 1st,8th,and 15th days.Rats in the pecking moxibustion group were treated with pecking moxibustion at Quchi(LI11),Dazhui(GV14),and Ashi points,and each point was treated with moxibustion for 15 min every day and a total of 3 courses of treatment,with 6 d as a course of treatment.After treatment,the capillary permeability,joint swelling,joint surface temperature,and plantar thermal pain threshold of the diseased joints in rats were observed,and the ultrastructural changes of synovial cells were observed by transmission electron microscopy. Results:The local swelling,surface temperature,and Evans blue(EB)leakage volume were significantly higher(P<0.05),the thermal pain threshold was significantly lower(P<0.05),and the synovial cell ultrastructure was obviously damaged in the affected joints in the model group compared with the blank group.The swelling degree,surface temperature,and EB leakage volume were significantly reduced(P<0.05),the thermal pain threshold was significantly increased(P<0.05),and the ultrastructural abnormalities of synovial cells were significantly improved in the diseased joints in the drug group and the pecking moxibustion group compared with the model group.The thermal pain threshold of rats in the pecking moxibustion group was significantly improved compared with the drug group(P<0.05). Conclusion:Pecking moxibustion obviously improves the pattern characteristics of local redness,swelling,heat,and pain in the diseased joints of rats with RA due to damp heat affecting bones/joints and effectively repairs the ultrastructure of the damaged synovium.It suggests that the pecking moxibustion intervention has a significant anti-inflammatory effect on early RA.