Objective To evaluate the ability of a synthetic PET/NIRF probe,named 18F-PSVue643,on the apoptosis detection in animal models with the anti-cancer drugs therapy,and compare the advantages and disadvantages between PET and NIRF.Methods Cell apoptosis was detected by MTT and flow cytometry in vitro.Established U87MG glioblastoma xenograft tumors in nude mice were treated with retinol and paclitaxel for nine days (for PET imaging) or eleven days (for NIRF imaging) continuously.The uptake values were recorded by ROI and expressed as ％ID/g.Results The apoptosis ratios in 10 and 100 nmol/L paclitaxel-treated groups were 7.4％ and 7.5％,respectively,and the apoptosis ratio of the control group was 4.3％.The apoptosis could be well detected by both NIRF and PET imaging during the whole process of treatment.However,the amount of probe for PET imaging was only a half of that for optical imaging to get the same apoptosis visualization.Conclusion 18F-PSVue643 is suitable for NIRF and PET imaging in detection of apoptosis,and it may be a promising agent for further clinical studies.

Objective To prepare dual?modality single?photon emission computed tomography (SPECT)?MRI molecular nanoprobes targeting HAb18G/CD147 expressed on breast cancer cell membranes and investigate the physicochemical and biological properties in vitro. Methods Superparamagnetic iron oxide nanoparticles (SPIOs) were prepared by one?pot reaction method as described. The single?chain antibody fragments HAb18F(ab')2 were conjugated to SPIOs via chemical method and then labeled with 125I using Iodogen method. The final 125I?SPIO?HAbF18(ab')2 nanoprobes were purified. SPIOs or 125I?HAb18F(ab')2 were used as control. We carried preliminary evaluation on their physicochemical properties and biological characteristics in vitro: transmission electron microscope (TEM) and dynamic light scattering (DLS) were used to measure these nanoparticle sizes and the hydrodynamic diameters. The MRI T2 transverse relaxation efficiency of these nanoprobes at different Fe2+concentrations were measured with 1.5 T clinical MR scanner. The 125I?SPIO?HAb18F(ab')2 and 125I?HAb18F(ab')2 radiochemical purity were measured by thin layer chromatography and the radio chemical yield was calculated. We also conducted stability tests in vitro and octanol/water partition coefficient experiments. Two breast tumor cell lines, MDA?MB?231 (HAb18G?overexpressing cells,experimental group) and MDA?MB?468 (control), were used for assessment of cells viability at different Fe2 + concentrations (1, 5, 10, 20, 40 μg/ml) by methyl thiazolyl tetrazolium assay. Specific binding experiments in vitro included two parts:magnetic resonance imaging and radionuclide tests, the above?mentioned breast cancer cell lines were incubated with 125I?SPIO?HAb18F(ab')2 nanoprobes respectively and took MDA?MB?231 cells which were not treated as blank group. First comparing the MR signal intensity differences among experimental group, the control group and blank group, then calculated the rate of MRI signal changes;Two breast tumor cell lines, MDA?MB?231 and MDA?MB?468 were incubated with 125I?SPIO?HAb18F(ab')2 nanoprobes too, then measured radioactivity counting byγcounter at different time and calculated the cell binding rates, and did statistical analysis by using one?way ANOVA. Results The SPIOs were fairly homogeneous with an average core size of (10.32±1.30) nm;the SPIO and 125I?SPIO?HAb18F(ab')2 hydrodynamic diameter of 44.80 and 52.64 nm, and MRI scanning showed that the transverse relaxation efficiency of SPIO and 125I?SPIO?HAb18F(ab')2 were 38.79 and 106.73 mM-1 · s-1, respectively. The radio chemical yield of 125I?SPIO?HAbF18(ab')2 and 125I?HAb18F(ab')2 were 41.90% and 85.50%, respectively. The radio chemical yield of the two groups were >95%, suggesting well stability in vitro. The lipo?hydro partition coefficient values were -0.99 ± 0.03 and-1.49 ± 0.08, respectively, which demonstrated that they were both water?soluble substances. Different Fe2+concentrations (1,5,10,20,40μg/ml) of 125I?SPIO?HAb18F(ab')2 on breast cancer cell lines MDA?MB?231 and MDA?MB?468 showed no significant inhibition of cell proliferation (F values were 0.78, 0.66; P values were 0.58, 0.66). The cell?specific binding experiment showed: MRI signal intensity values on experimental group, the control group and the blank group were (1 670 ± 5), (1 930 ± 8), (2 349 ± 14), respectively, significant differences existed among these groups (F=4 408.48,P=0.000), the rate of signal intensity change of experimental group and the control group were 28.87%,17.78%. SPECT:MDA?MB?231 could uptake 125I?SPIO?HAb18F(ab')2, the cell binding rates were (6.52 ± 0.60)% and (10.52 ± 2.04)% in 20 min and 4 h, respectively.Conclusions Our results suggested that the dual?modality SPECT?MRI nanoprobes 125I?SPIO?HAb18F(ab')2 were prepared successfully with good physicochemical properties and biological characteristics in vitro. These dual?modality molecular imaging nano?probes may have potential to improvearly detection and diagnosis of HAb18G/CD147?expressing cancers and to facilitate the development of HAb18G/CD147?directed interventions.

Objective To study the effect of Buyang-Huanwu decoction on incidence of macular edema, visual acuity and serum VEGF, IL-6 levels after operation in diabetic cataract patients. Methods A total of 46 post-operative patients with diabetic cataract were randomly divided into two groups. The control group was treated with pranoprofen eye drops combined with tobramycin dexamethasone eye drops, and the study group was treated with Buyang-Huanwu decoction combined with Salviae Miltiorrhizae and Ligustrazine Hydrochloride injection based on the control group. The central subfield mean thickness (CSMT) at baseline before operation and 1d, 1w, 4w, 8w, 12w after operation, corrected vision at 0w, 1w, 6w, 12w, serum levels of vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) before and after treatment of two groups were observed, and the incidence of macular edema was analysed. Results The CSMT of study group at 8 w (226.35 ± 9.88 μm vs. 214.74 ± 9.27 μm, t=2.118) and 12 w (233.57 ± 11.25 μm vs. 213.62 ± 10.13 μm, t=2.258) after operation were significantly lower than those of control group (P＜0.05). The best corrected visual acuity of study group at 6 w (4.61 ± 0.32 vs. 4.82 ± 0.25, t=2.0460 and 12 w (4.57 ± 0.29 vs. 4.79 ± 0.24, t=2.180) after operation were significantly higher than those of control group (P＜0.05). The levels of VEGF (174.31 ± 45.63 pg/ml vs. 83.65 ± 24.47 pg/ml, t=4.145) and IL-6 (27.25 ± 9.21 pg/ml vs. 15.24 ± 3.14 pg/ml, t=3.829) of study group were significantly lower than those of control group (P＜0.05). The incidence of macular edema at 8 w and 12 w after operation in study group were respectively 0.0% and 2.9% (1/35), and the control group were 16.2% (6/37) and 21.6% (8/37). The difference was statistically significant (P＜0.05). Conclusions The Buyang-Huanwu decoction in treatement of post-operation patients with diabetic cataract could reduce the incidence of macular edema and improve the visual acuity.

Objective To evaluate the effect of Xiaoyaosan and Shengmaiyin combined with conventional western medicine for xerophthalmia.Methods A total of 76 xerophthalmia patients who met the criteria were divided into two groups according to random number table,with 38 cases in each group.The control group was treated with routine western medicine,while the observation group was treated with Xiaoyaosan combined with Shengmaiyin on the basis of the control group.Both groups were treated for 4 weeks.Before and after treatment,the symptoms were scored,and the tear film stability was evaluated according to the results of fluorescent test (FL),breakup time of tear film (BUT) and schirmer Ⅰ test (SIT).The contents of IL-1β,IL-6 and TNF-α in tear were detected by ELISA,and the clinical efficacy was evaluated.Results The total effective rate was 92.9％ (65/70) in the observation group and 76.4％ (55/72) in the control group,and there was significant difference between the two groups (Z=-2.991,P=0.002).After treatment,the SIT (6.9 ± 0.8 mm vs.4.3 ± 0.5 mm,t=3.751),BUT (10.5 ± 1.6 s vs.6.4 ± 0.8 s,t=4.228) in the observation group were significantly higher than those in the control group (P＜0.05).The FL score (0.9 ± 0.1 vs.1.4 ± 0.2,t=3.208) in the observation group was significantly lower than that of the control group (P＜0.05).The scores of aningeresting,foreign body sensation,visual fatigue,photophobia and visual blur in the observation group were significantly lower than those in the control group (t were 3.559,4.015,4.119,3.983,4.120,all Ps＜0.05).The levels of IL-1β,IL-6 and TNF-α in tear were significantly lower than those in control group (t were 11.887,8.028,8.112,all Ps＜0.001).Conclusions The Xiaoyaosan combined with Shengmaiyin can improve tear film stability,relieve local ocular surface inflammation and improve clinical symptoms in patients with xerophthalmia.

Objective To investigate the effect of viral myocarditis serum exosomal miR-320 on apoptosis of cardiomyocytes and its mechanism. Methods The model of viral myocarditis mice was established by intraperitoneal injection of Coxsackie virus B3. Serum exosomes were extracted by serum exosome extraction kit and co-cultured with cardiomyocytes. The uptake of exosomes by cardiomyocytes was detected by laser confocal microscopy. Cardiomyocytes were transfected with miR-320 inhibitor or mimic, and the expression level of miR-320 was detected by real-time quantitative PCR. Flow cytometry was used to detect cardiomyocyte apoptosis rate, and the expression levels of B cell lymphoma 2 (Bcl2) and Bcl2-related X protein (BAX) were tested by Western blot analysis. The prediction of miR-320 target genes and GO and KEGG enrichment analysis were tested by online database. The relationship between miR-320 and its target gene phosphoinositide-3-kinase regulatory subunit 1(Pik3r1) was examined by luciferase reporter gene. The effect of miR-320 on AKT/mTOR pathway protein was detected by Western blot analysis. Results Viral myocarditis serum exosomes promoted cardiomyocyte apoptosis, and increased the level of BAX while the level of Bcl2 was decreased. miR-320 was significantly up-regulated in myocardial tissue of viral myocarditis mice, and both pri-miR-320 and mature of miR-320 were up-regulated greatly in cardiomyocytes. The level of miR-320 in cardiomyocytes treated with viral myocarditis serum exosomes was significantly up-regulated, while transfection of miR-320 inhibitor counteracted miR-320 overexpression and reduced apoptosis rate caused by exosomes. Pik3r1 is the target gene of miR-320, and its overexpression reversed cardiomyocyte apoptosis induced by miR-320 up-regulation. The overexpression of miR-320 inhibited AKT/mTOR pathway activation. Conclusion Viral myocarditis serum exosome-derived miR-320 promotes apoptosis of mouse cardiomyocytes by inhibiting AKT/mTOR pathway by targeting Pik3r1.
Mice ; Animals ; Myocytes, Cardiac ; Phosphatidylinositol 3-Kinase/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Myocarditis/pathology* ; Exosomes/metabolism* ; bcl-2-Associated X Protein/metabolism* ; MicroRNAs/metabolism* ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis/genetics*