Objective To investigate the anti-fatigue and anti-hypoxia effect of fermented Paecilomyces hepiali myceli-um (PHM) and to explore its mechanism.Methods Seventy-two naive Kunming male mice were randomly divided into Home, vehicle, modafinil( Mode) , low-dose PHM, moderate-dose PHM, and high-dose PHM groups.All mice were ad-ministrated with the drug or vehicle twice a day during a 14-day period.Except for the Home group, each groups was forced into the wheel fatigue device to receive a climb-run training at the time points of 40 min, 8 h, 24 h and 48 h, and to re-ceive an exhaustive training at the time point of 72 h after the last administration.The number of electric shocks was recor-ded during each training and the levels of muscle glycogen, liver glycogen, serum urea nitrogen, serum lactic acid, serum creatine kinase and blood ATP were detected after the exhaustive training using commercially available kits.In addition, 96 naive Kunming male mice were randomly and equally divided into the vehicle, low-dose, moderate-dose and high-dose PHM groups.Twelve mice in each group were chosen for the normal pressure anti-hypoxia experiment and the other 12 mice were used for the NaNO3 poisoning experiment.Results Compared with the vehicle groups, the PHM group showed a sig-nificant decrease in the number of electric shocks, improvement in biochemical parameters associated with fatigue, and an increased survival time in the anti-hypoxia and the NaNO3 toxicity tests.Conclusion PHM is potentially an effective alter-native for wild Cordyceps in the treatment of fatigue and hypoxia.

Purpose To investigate the value of 3.0T MR T1ρquantitative analysis in early detection of disc degeneration. Materials and Methods Conventional T2WI and T1ρwere collected in 35 healthy volunteers (male 16, female 19) on 3.0T MRI, and the classification of disc nucleus was performed by Pfirrmann classification. T1ρvalue of nucleus pulposus was measured and analyzed for the relationship among Pfirrmann classification, segmental and gender. Results There was a significant negative correlation between T1ρvalue and Pfirrmann grade (r=-0.542, P<0.001). T1ρvalue in L5/S1 segment (94.80±26.60) ms was significantly lower than that in L2/L3 (117.18±25.64) ms and L3/L4 (115.52±28.53) ms (P<0.01), and there was no significant difference among other segments (P>0.05);there was no gender differences among the T1ρvalue of each segment (t=0.006, 0.042, 0.797, 1.022, 0.038, P>0.05). Conclusion T1ρ value is closely related to the degree of disc degeneration, and T1ρimaging can be used as an objective, sensitive and effective tool for early detection of disc degeneration.

Objective To evaluate the possibility and scan the protocol of diffusion tensor imaging(DTI) in rabbit VX2 liver tumor models.Methods MRI and DTI images were obtained from 16 rabbit VX2 liver tumor models and 4 rabbit control liver models.Average diffusion coefficient(ADC),fractional anisotropy(FA) of tumors and control livers were estimated and compared with the changing rules(b value=100,600,1000 mm2/s).Results ADC of tumors had a descending trend and FA of tumors had an increasing trend compared with control livers.The fluctuations of ADC and FA of tumors had a descending trend(P

ObjectiveTo evaluate the characteristics of MRI of common hepatic lesions by using measurement of T1,T2 and ADC on the MRI and DWI. MethodsMRI and DWI were obtained in 100 patients with common hepatic lesions, and T1,T2,ADCs and the ratios of the ADC of lesion/liver were estimated. (Results)Average T1,T2 and ADCs of hepatic lesions were as follows: hepatocellular carcinoma, (843.92167.75) ms , (73.9014.21) ms and(0.990.26)10~(-3)mm~2/s respectively; hepatic (metastasis), (946.89186.13) ms, (76.3919.76) ms and(1.170.32)10~(-3)mm~2/s (respectively); hepatic cavernous hemangioma, (1102.33213.12)ms, (142.3228.51) ms and (1.810.42)10~(-3)mm~2/s respectively; hepatic cyst, (1516.32617.84)ms ,(247.33112.52)ms and (3.110.38)10~(-3)mm~2/s respectively. The ratio of the ADC of lesion/liver:(hepatocellular) carcinoma was 0.890.17, and significantly lower than the ratio of hepatic metastases((1.27)0.21, P

Objective:To investigate the expression of microRNA-324-5p (miR-324-5p) and transcription factor forkhead box C1 (FOXC1) in glioma and their relationship with the prognosis of patients.Methods:From March 2012 to March 2015, a total of 72 cases of glioma tissues were collected from glioma patients who were admitted to Chongqing Hygeia Tumor Hospital and the People's Hospital of Nanchuan in Chongqing, and 28 cases of normal human brain tissues resected in craniocerebral surgery were also collected. The expressions of miR-324-5p and FOXC1 mRNA were detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression of FOXC1 protein was detected by immunohistochemistry. Pearson method was used to analyze the correlation between the expressions of miR-324-5p and FOXC1 in glioma tissues; Kaplan-Meier method was used to analyze the survival of patients with glioma; Cox regression analysis was used to analyze the risk factors affecting the prognosis of patients with glioma.Results:FOXC1 protein was mainly located in the cytoplasm of glioma, and its positive expression rate in glioma tissues was 81.94% (59/72), which was significantly higher than that in normal brain tissues [17.86% (5/28)], and the difference was statistically significant ( χ2 = 35.938, P<0.01). Compared with normal brain tissues, the expression of miR-324-5p was down-regulated in glioma tissues (0.62±0.19 vs. 0.98±0.02, t = 9.974, P < 0.05), and the expression of FOXC1 mRNA was up-regulated (1.41±0.29 vs. 0.99±0.02, t = 7.633, P < 0.05). The expressions of miR-324-5p and FOXC1 protein were correlated with the number of primary lesions, differentiation degree, TNM stage and lymph node metastasis of glioma (all P<0.05). Pearson analysis showed that the expressions of miR-324-5p and FOXC1 mRNA were negatively correlated ( r = -0.550, P<0.01). The 5-year overall survival rate of patients in miR-324-5p high-expression group was significantly higher than that of patients in miR-324-5p low-expression group (45.71% vs. 24.33%, χ2 = 6.531, P = 0.011), and the 5-year overall survival rate of patients in FOXC1 protein high-expression group was significantly lower than that of patients in FOXC1 protein low-expression group (30.41% vs. 42.34%, χ2 = 3.631, P = 0.047). Multivariate Cox regression analysis showed that low differentiation, TNM stage Ⅲ-Ⅳ, lymph node metastasis, low expression of miR-324-5p and high expression of FOXC1 protein were independent risk factors for prognosis of glioma patients (all P < 0.05). Conclusions:The expression of miR-324-5p is low and the expression of FOXC1 is high in glioma. They may be involved in the regulation of tumor differentiation and metastasis, and related to the poor prognosis of patients. They may be potential therapeutic targets for glioma.

OBJECTIVETo prepare the quercetin self-emulsified formulation and evaluate its quality.

METHODThe quercetin self-emulsified formulation was optimized based on the quercetin solubility in different oils, and the self-microemulsified efficiency of various combinations of emulsifier and co-emulsifier evaluated using the pseudo-ternary phase diagram. The microemulsion of morphology, size and zeta potential were examined. The quercetin of solubility in self-emulsified system was tested and the formulation stability was investigated by accelerated experiment.

RESULTThe blank self-emulsified system was composed of ethyl oleate/Cremophor EUL/butanol with weight ratio of 10: 54: 36. After being dilutied with water, the morphology of microemulsion was homogeneous small spherical drops observed under the electro-microscopy. The particle size and the zeta potential were 16.3 +/- 4.6 nm and 2.1 +/- 0.8 mV, respectively. The solubility of quercetin in self-emulsifing system was (62.42 +/- 0.11) mg x mL(-1), increased 2 229 folds compared with that of in water. The quality of quercetin self-emulsified formulation was stable during the 3 months storage at 40 degrees C.

CONCLUSIONThe solubility of quercetin is significantly increased in self-emulsified system and the formulation is stable and easy to prepare.

Antioxidants ; administration & dosage ; chemistry ; Butanols ; chemistry ; Chromatography, High Pressure Liquid ; Drug Delivery Systems ; Drug Stability ; Emulsions ; Glycerol ; analogs & derivatives ; chemistry ; Oleic Acids ; chemistry ; Particle Size ; Quercetin ; administration & dosage ; chemistry ; Solubility ; Solvents ; Technology, Pharmaceutical ; methods

AIMTo investigate factors affecting the properties of antisense oligodeoxy nucleotides (ASON)-liposomes complex and their cellular uptake.

METHODSThree types of blank liposomes were prepared by reverse-phase evaporation vesicles, and the complex were obtained through physical absorption. The light microscope was used to observe morphology characteristics of the complex. Drug loading capacity was analyzed by agarose gel electrophoresis. The transfected cell percentage and means fluorescence intensity were determined by flow cytometric analysis using M3 myeloma cell as a model.

RESULTSThe neutral liposome showed no aggregation while the cationic liposomes appeared some different extent aggregation in different medium when associated antisense oligodeoxynucleotides. The drug loading capacity depended on the ratio of +/- and the cationic charge density on the lipid membrane. The two kinds of cationic liposomes appeared different principles of loading ASON. As far as cellular uptake, The neutral liposomes showed no improvement of cellular uptake of ASON. However, the cationic liposomes were shown to enhance the cellular uptake of ASON if the appropriate +/- charge ratio was used. The optimal cellular uptake was achieved when +/- charge ratio was at 0.5:1 and 1:1 for SA-I liposome and SA-II liposomes, respectively.

CONCLUSIONThe cationic liposomes improved the loading capacity and cell uptake of antisense oligodeoxynucleotides, which was determined by +/- charge ratio and charge density.

Amines ; metabolism ; Biological Transport ; Drug Carriers ; Drug Delivery Systems ; Humans ; Liposomes ; pharmacokinetics ; Multiple Myeloma ; Oligodeoxyribonucleotides, Antisense ; administration & dosage ; pharmacokinetics ; Random Allocation ; Tumor Cells, Cultured ; metabolism

AIMTo investigate the possibility of polymethacrylate nanoparticles (NP) for antisense oligodeoxynucleotides delivery system.

METHODSThe nanoparticles were prepared by evaporating ethenol solution containing Eudragit RL100 or RS100, and then mixtured with oligonucleotides. The morphology and size were investigated by a transmission electron microscope and Mastersizer particle characterization systems, and the cytotoxicity was evaluated by Trypan Blue staining and hemolysis test. The flow cytometer was used to determine the uptake of fluorescence-labelled oligodeoxynucleotides.

RESULTSThe morphology of nanoparticles showed spherical and orderly, the average diameter was about 127 nm, and almost the antisense oligodeoxynucleotides (ODN) were loaded when NP: ODN was 6.6. The uptake of ODN was significantly increased when loaded by nanoparticles, which well depended on the nanoparticles concentration. Meanwhile, slightly cytotoxicity was observed when high dose of nanoparticles was used.

CONCLUSIONThe polymethacrylate nanoparticles appeared to be a promising vehicle for gene delivery.

Acrylic Resins ; chemistry ; toxicity ; Animals ; Drug Carriers ; Drug Delivery Systems ; Hemolysis ; drug effects ; Nanotechnology ; Oligonucleotides, Antisense ; administration & dosage ; Particle Size ; Technology, Pharmaceutical ; methods

AIMTo survey the uptake behavior and subcellular distribution of antisense oligodeoxynucleotide polymethacrylate submicroparticles (AS-ODN-SMP) and infer its mechanism in MGC cell lines.

METHODSMGC cells were incubated at certain concentration of AS-ODN-SMP or AS-ODN for 8 h at 4 degrees C or 37 degrees C. Then the fluorescence oligodeoxynucleotide- labeled cells were counted by flow cytometer and the intracellular fluorescence intensity was determined after incubated with chloroquine for 2 h.

RESULTSCellular uptake of oligodeoxynucleotides was significantly increased following application of AS-ODN-SMP and total intracellular fluorescence intensity was enhanced by 683 folds with the vehicle concentration of 20 microg x mL(-1). AS-ODN-SMP entranced to cells profoundly with temperature-dependent manner. Rare cells took on fluorescence when incubated at 4 degrees C, while 37 degrees C they were significantly increased. But the intracellular fluorescence intensity appeared same level in present or absent of chloroquine.

CONCLUSIONWith the help of polyacrylate submicroparticles, oligonucleotides efficiently entranced the cells via endocytosis and could successfully escape the degradation in lysosome.

Animals ; Cell Line ; Drug Carriers ; Drug Delivery Systems ; Endocytosis ; drug effects ; Giant Cells ; cytology ; Lysosomes ; metabolism ; Nanoparticles ; Oligodeoxyribonucleotides, Antisense ; administration & dosage ; pharmacokinetics ; Particle Size ; Polymethacrylic Acids ; chemistry ; pharmacology ; Temperature

OBJECTIVE: To investigate a nwe, simple technique for preparation of interferon-alpha-liposomes, which may be suitable for industrial use. METHODS The uniform design coupled with computerized optimization was utilized to screen the formulation and preparation procedure of interferon-alpha-liposomes. Pro-liposomes were prepared by the powder bed grinding method and combined with interferon-alpha-solution to form interferon-alpha-liposomes. Liposome size was determined by the particle size analyzer. Free interferon-alpha and interferon-alpha-liposome were separated by gel filtration. Then the recovered activity of interferon-alpha was analyzed by enzyme-linked immunosorbent assay. RESULTS The result demonstrated that the best interferon-alpha-liposome formulation was as follows: the protectant was sorbitol; weight ratio of protectant to lipid was 5:1; weight ratio of octadecytamin to lipid was 1:9; weight ratio of sobey phosphatidylcholine to cholesterol was 9:1 respectively. Interferon-alpha-liposome size determined by the particle size analyzer was 80.8+/-36 nm and the encapsulation efficiency was 59.0+/-3.3%. CONCLUSION The powder bed grinding method can be used to prepare pro-liposomes which can be easily combined with interferon-alpha-solution to form interferon-alpha-liposomes.