The preparation method, serum stability, efficiency of cellular uptake and apoptosis induction of the cell penetrating peptide TAT and cleavable PEG co-modified liposomes loaded with paclitaxel (C-TAT-Lipo) were investigated. The best preparation procedure was performed by orthogonal test based on single factor screening method. First, the paclitaxel (PTX)-loaded liposomes were prepared by filming-rehydration method, evaluated with entrapment efficiency and polydispersity index. The morphology of C-TAT-Lipo was characterized by transmission electron microscopy. Turbidity variations were monitored in the presence of fetal bovine serum (FBS) to evaluate the serum stability of the liposomes developed here. Next, the efficiency of cellular uptake of different Rho-PE-labeled liposomes on B16F1 cells in vitro was evaluated by confocal laser scanning microscopy (CLSM) and flow cytometry. The quantitative analysis of apoptosis induced by different PTX-loaded liposomes was performed by Annexin V-FITC/PI double staining. The optimal formulation was as follows: Chol : lipid: 1 : 8 (molar ratio); drug : lipid: 1 : 40 (mass ratio); lipid concentration: 3 mmol x L(-1); temperature of hydration: 25 degrees C. The mean size and polydispersity index of C-TAT-Lipo were about (97.97 +/- 3.68) nm and 0.196 +/- 0.037, the zeta potential was (-0.89 +/- 0.45) mV, the entrapment efficiency of paclitaxel was (90.16 +/- 1.53)%. The particle sizes did not exhibit significant variations in 50% FBS over 24 h at 37 degrees C. The efficiency of cellular uptake of the C-TAT-Lipo increased 1.40 fold following the cleavage of PEG. Apoptosis analysis showed 59.3% increase of the apoptosis and necrosis profile of C-TAT-Lipo after the detachment of PEG shells, which was markedly higher than that of N-TAT-LP with or without glutathione and SL, respectively. The results indicate that the C-TAT-Lipo is successfully prepared by filming-rehydration method and shows significant antitumor activities.
Animals ; Annexin A5 ; Apoptosis ; Cell Line, Tumor ; Cell-Penetrating Peptides ; pharmacology ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; Liposomes ; chemistry ; Melanoma, Experimental ; Mice ; Microscopy, Confocal ; Paclitaxel ; pharmacology ; Particle Size ; Polyethylene Glycols ; chemistry

Bilirubin ; Coronary Artery Disease ; Humans

Objective To investigate the impact of health education and standard treatments on the life quality of asthmatic children.Methods We conducted clinic treatments of 110 cases of asthmatic children lasting for 6 months,which were divided into standard treatment and health education group with 60 cases and group without standard treatment and health education with 50 cases.We emphasized the education management on cases in experimental group,which strictly obeyed the Global Initiative for Asthma (GINA) to formulate classified treatments as well as regular follow-up.Control group received the same treatments as experimental group yet without emphasizing health education about asthma on parents,which was accompanied with irregular follow-up.We compared the before-treatment and after-treatment life-quality and lung functions of asthmatic children.Moreover,we used Pediatric Asthma Quality of Life Questionnaire (PAQLQ) to evaluate the before-treatment and after-treatment life quality of asthmatic children.Results After follow-up lasting for 6 months,the treatments of experiment group had obvious improvements (P ＜0.01).The improvements of peak expiratory flow (PEF) of lung function test and timed vital capacity of the first second (FEV1) of experimental group were also better than the improvements of these two metrics of control group (P ＜ 0.01).In addition,after 6 months,in both all dimensions of PAQLQ and total score,the differences between before and after treatment of experimental group were significant (P ＜ 0.01,P ＜ 0.05) ; and the differences between experimental group and control group were also significant in both all dimensions of PAQLQ score (12.4 ±2.1) and total score comparing to the control group (8.1 ±2.3),and the differences are also significant (t =2.5,P ＜ 0.01).Conclusions Health education and standard treatments significantly improved the life quality of asthmatic children with a good compliance of treatment and a low recurrence rate,which is worth of popularizing.

AIM: To express the DT389-hbFGF (389 amino acid residues of the N-terminus of diphtheria toxin (human basic fibroblast growth factor) fusion protein for potential targeting therapy towards posterior capsule opacification (PCO) after cataract surgery.METHODS: The DNA of inactivated diphtheria bacillus and RNA of 12-week fetal brain cortex were extracted, respectively. The fragments of truncated diphtheria toxin (containing 389 amino acids of N-terminus, DT389) )and full-length human basic fibroblast growth factor(hbFGF) sequence (encoding 18kDa protein) were amplified by PCR. The two fragments were inserted into pGEX-4T-1 prokaryotic expression vector to obtain pGEX-DT389-hbFGF prokaryotic expression plasmid. After sequence analysis, the expressing plasmid was transformed into Escherichia Coli BL21 strain and expression was induced under IPTG. The expressed fusion protein was purified and identified.RESULTS: The gene fragments encoding DT389 and hbFGF were amplified and their gene sequences were confirmed. Hybrid gene expression plasmid pGEX-DT389 (hbFGF was constructed. The fusion protein DT389-hbFGF was expressed and purified.CONCLUSIONS: The successful cloning and expression of DT389-hbFGF immunotoxin provides a foundation for targeting therapy towards posterior capsule opacification.

BACKGROUND:Ankylosing spondylitis is an autoimmune disease involved in chronic systemic inflammation. Tumor necrosis factor and interleukin-6 levels increased in patients with ankylosing spondylitis. Inflammatory factors such as tumor necrosis factor and interleukin-6 can suppress CD36 expression in monocytes. OBJECTIVE: To analyze the correlation between CD36 expression in monocytes and ankylosing spondylitis. METHODS:A total of 84 newly diagnosed ankylosing spondylitis patients and 111 healthy individuals were included in this study. CD36 expressions in monocytes in ankylosing spondylitis patients and healthy individuals were tested using flow cytometer; meanwhile, biochemistry, immunology, routine blood examination and related inflammatory markers were determined between the two groups. RESULTS AND CONCLUSION:Results of baseline data in both groups demonstrated that CD36 fluorescence intensity in monocytes was significantly lower in patients with ankylosing spondylitis compared with healthy controls (P < 0.01). CD36 fluorescence intensity in monocytes was negatively correlated with C-reactive protein, erythrocyte sedimentation, interleukin-6 and tumor necrosis factor. In addition, CD36 fluorescence intensity in monocytes was negatively correlated with BASDAI score. Logistic regression analysis showed that erythrocyte sedimentation, interleukin-6, tumor necrosis factor and CD36 fluorescence intensity in monocytes were associated with ankylosing spondylitis, and risk factors for ankylosing spondylitis (P < 0.05). These findings confirm that inflammatory cytokine in patients with ankylosing spondylitis weakened the expression of CD36 in monocytes. There was a remarkable association between low expression of CD36 expression in monocytes and ankylosing spondylitis. CD36 expression of monocytes clinicaly may be considered to be an effective indicator to evaluate inflammation and disease activity in patients with ankylosing spondylitis.

Verification of scientific literature novelty assessment is the final step in writing the literature novelty assessment report and is closely related with its academic level.The key points in verifying novelty assessment of medical literature were thus analyzed with examples in this paper, including the scientific and technical points, literature novelty assessment points, retrieval terms, retrieval strategies, and conclusion of literature novelty assessment.

Objective To observe the baseline characteristics and visual outcomes after two years follow-up of exudative age-related macular degeneration (AMD) patients treated with ranibizumb. Methods Forty-four eyes of 44 patients with exudative AMD were enrolled into this retrospective study, 19 were men and 25 were women. The mean age was 78 years (range 64–92 years). All patients were underwent best corrected visual acuity (BCVA, Early Treatment of Diabetic Retinopathy Study), fundus color photography, fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA) and optical coherence tomography (OCT). The mean BCVA was (50.36±14.43) letters, the mean central foveal thickness (CFT) was (291.95± 82.19)μm, and the fluorescence leakage area of choroidal neovascularization (CNV) was (7.61±5.84) mm2. All patients received three initial intravitreous injection of ranibizumb (IVR) and were retreated with monthly IVR when needed. The mean follow up time was 25.6 months (range 24–29 months). On 1, 2, 3, 6, 12, 18 and 24 months after treatment, BCVA and OCT were repeated. On 3, 6, 12, 18 and 24 months after treatment, FFA and ICGA were repeated. The change of BCVA, CFT and fluorescence leakage area of CNV were observed. The association of baseline characteristics and two year visual outcomes were analyzed. Results On 1, 2, 3, 6, 12, 18 and 24 months after treatment, the BCVA were improved significantly (t=?1.89,?3.51,?4.61,?4.04,?5.77,?4.69;P<0.05), the CFT were decreased significantly (t=1.51, 2.30, 3.40, 3.28, 3.54, 3.88, 3.73;P<0.05). On 3, 6, 12, 18 and 24 months after treatment, the fluorescence leakage area of CNV were reduced significantly (t=2.12, 2.90, 3.51, 4.12, 4.06;P<0.05). The lower baseline BCVA, the more improved after treatment. The BCVA improvement degree has a negative relationship with baseline BCVA and fluorescence leakage area of CNV (r=0.505,?0.550;P<0.05), but no correlation with baseline CFT (r=0.210, P>0.05). Conclusion Two year visual outcomes of exudative AMD patients treated with ranibizumb is negative correlated with baseline BCVA and fluorescence leakage area of CNV, but not correlated with baseline CFT.

[Objective]To study the effects of different bionic microstructure of artificial bones on the growth and osteogenesis characteristics of dog bone marrow stromal cells(BMSCs).[Method]The artificial bones were divided into 4 groups in the experiments:perpendicular structure(experimental group,A),concentric structure(experimental group,B),CPC/ rhBMP-2(calcimrt phosphate cement composited with recombinant human bone morphogenetic protein 2)(control group,C)and pure CPC(control group,D).Dog bone marrow mononuclear cells were isolated from the bone marrow by gradient centrifugation and then cultured in non-induced condition.BMSCs at 3~(rd) passage were harvested and inoculated onto the artificial bones of the 4 groups.The implanted cells and artificial bones were retrieved and observed at different times with fluorescence microscope,scanning electron microscope.The proliferation of BMSCs was evaluated by MTT method,and the osteogenic differentiation was evaluated by alkaline phosphatase(ALP)activity.[Result]The cultured BMSCs adhered well to the surface of all artificial bones and it was better to A and B groups than to C and D groups.By MTT method,the A values of group A,B,C were significantly higher compared with group D(all P0.05);The ALP activities of BMSCs in group A,B,C and D all increased significantly from the 4~(th) day to the 7~(th) day(P0.05).[Conclusion]The bionic microstrueture artificial bones are satisfactory carriers of BMP and possess good cellular biocompatibility with dog bone marrow stromal cells and have the ability to enhance the osteogenic differentiation of BMSCs.

[Objective] To establish a method to seperate rat mesenchymal stem cells(rMSCs) from rat marrow, culture rMSCs in vitro, and study its biological characteristics. [Methods] rMSCs were seperated from rat marrow with wall-sticking method and expanding cultured in vitro. To draw the growth curves of cells at 1th, 3th, 5th passage, detect the surface antigens with flow cytometry. To evaluate the effect of fetal calf serum with different concentrations to rMSCs. [Results] There were approximately (5～6)?105 cells after primary culture and over (2~3)?108 rMSCs at 5th passage. The appearance of rMSCs is like shape of spear and its growth curves are like shape of S. The rMSCs were positive for CD90 and negative for CD45. [Conclusions] Cells seperated and cultured by the method established in this experiment were rMSCs. They have the biological characteristics of rMSCs, and are good resources for study in tissue engineering.

Animals ; Brain ; metabolism ; Brain Ischemia ; blood ; enzymology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Nitric Oxide ; blood ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism