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

Objective: To establish the methods for extraction and isolation of punicalagin from pomegranate peel, and further study the purification and quantification of punicalagin. Methods: Using an ultrasonic-assisted extraction method, punicalagin in pomegranate peel was extracted at room temperature by 50% ethanol with 20-fold volume of raw material. The content of punicalagin in the crude extract was determined by HPLC. To optimize the purification process of punicalagin, static adsorption and desorption experi-ments were employed to study five kinds of macroporous adsorbent resins (D101, A8-8, NKA-9, HPD-100 and HPD-500) for the one with the highest purification efficacy of punicalagin. In addition, the technical parameters of the macroporous adsorbent resin were opti-mized to obtain punicalagin with higher purity. Punicalagin was further separated and purified by using a reverse phase MCI GEL CHP20P column. Results:HPD500 resin showed the best ability to absorb and separate punicalagin in among five kinds of macro-porous adsorbent resins. The best technical parameters were as follows:the mass concentration of sample solution was 15 mg·ml-1 , the loading amount was 2BV, the pH was 2 and the eluting solvent was 8BV of 30% ethanol. With the best process as described a-bove, the content of punicalagin extracted from pomegranate peel increased from 10. 3% to 30. 7%. The obtained punicalagin could be further purified to 61. 3% from 30% in ethanol eluate by the reverse phase MCI GEL CHP20P column. Conclusion:HPD500 resin is the most effective in the purification of punicalagin from pomegranate peel, and the content of punicalagin can be dramatically increased after the purification by a reverse phase MCI GEL CHP20P column. The optimized process shows good reproducibility and stability.

Objective To study the correlation between rotavirus nonstructural protein 1(NSP1) and species restriction of the virus and explore the role of NSP1 in the species restriction.Methods NSP1 C-terminal sequences,its whole amino acid sequences from UniProt Database,and its 3' noncoding sequences from GenBank were collected,and intraspecies and interspecies sequence alignment and phylogenetic tree analysis were performed by DNAStar(Lasergene)7.1.Results The diversity of NSP1 between species and rotavirus species restriction hosted a high degree of consistency.NSP1 C-terminal sequence,NSP1 gene 3' noncoding sequence and NSP1 whole sequence shared a high intraspecies identity,which was higher than the identity between species(P

Salvianolic acids are the main water-soluble active compounds of Salvia miltiorrhiza and have been widely used for the treatment of cardiovascular diseases. Based on the latest studies in China and abroad, we summarize the pharmacological effects and mechanism of salvianolic acids on ischemic heart disease by describing how salvianolic acid A and salvianolic acid B protect the vascular endothelium, relax coronary arteries, promote angiogenesis and anti-platelet aggregation, inhibit the inflammatory response, anti-cell apoptosis, and scavenge free radicals. This review provides a theoretical basis for further research on the effects of salvianolic acids on ischemic heart disease and their potential for drug development.

Ischemic heart disease (IHD), which has been considered to be exclusively caused by stenosis or occlusion of coronary artery, is a significant cause of morbidity and mortality worldwide. Mitochondrial dysfunction is the main pathological basis of ischemic heart disease and reperfusion injury, and moderate mitochondrial autophagy can selectively remove damage proteins and organelles to maintain intracellular homeostasis, so mitochondrial autophagy is important for maintaining the homeostasis of cardiomyocytes. Natural drugs from plants are widely used in ischemic heart disease. In recent years, more and more natural drugs have been proven to alleviate myocardial cell damage after ischemia/reperfusion through mitochondrial autophagy. This paper reviews the research progress of natural drugs from plants medicines regulating mitochondrial autophagy in the treatment of ischemia heart disease.

OBJECTIVETo explore metabolite profiling changes in serum of rats with pi-qi deficiency syndrome (PQDS) and pi-yang deficiency syndrome (PYDS) based on liquid chromatograph-mass spectrometer (LC-MS) technique, and to explore the essence of Pi-deficiency syndrome (PDS) from small molecule metabolite level.

METHODSTotally 21 male SD rats of SPF grade were randomly divided into three groups, the normal control group, the PQDS group, and the PYDS group, 7 in each group. Rats in the PQDS group overate for 1 day and fasted for 2 days. They drank freely and then swam to be exhausted in water at 35 degrees C - 37 degrees C for 15 successive days. The PYDS model was established by the same method for PQDS rats plus drenching 20% Folium sennae water extract (2 mL/100 g), once in the morning and once in the evening for one successive week. After modeling, models were evaluated according to rat general state, changes in body weight and rectal temperature. Serum metabonomic profiles were detected using LC-MS technique. Difference in inter-group metabolite spectrograms was analyzed using orthogonal partial least squares discriminant analysis (OPLS-DA). Potential biomarkers related to syndrome types in rat serum were selected via the parameter of variable importance in the projection (VIP).

RESULTSThe weight of rats in the PQDS group and the PYDS group decreased more significantly after modeling. The difference in prepost weight was statistically significant from that of the normal control group (P < 0.01). It was more obviously lowered in the PYDS group than in the PQDS group (P < 0.05). Compared with the normal control group, the rectal temperature of rats in the PYDS group and the PQDS group decreased (P < 0.05, P < 0.01). It decresed more in the PYDS group than in the PQDS group (P < 0.05, P < 0.01). Compared with the normal control group, levels of PC(19:0)/PE(22:0), PC(17:0)/PE(20:0), capric acid, oleic acid, stearic acid, succinic acid, fumaric acid, malic acid, glucose increased; arachidonic acid, linolenic acid, lauric acid, androsterone, 4-heptanone, dihydroxyacetonephosphate (DHAP) (6:0), and uridine decreased in the PYDS group and the PQDS group. Compared with the PQDS group, levels of PC(22:1), PC (22:6), PE (18:0)/PC (15:0), retinol, and deoxycytidine increased significantly in the PYDS group; PC (18:1), PC(19 :3), PC (20:3), PC (17:0)/PE (20:0), PC (19:1)/PE (22:1), PC (19:0)/PE (22:0), PC (17:1)/PE (20: 1), PC (16:1)/PE (19:1), cholic acid, hippuric acid, furoic acid, undecanedicarboxylic acid, palmitoleic acid, hydroxy stearic acid, eicosatrienoic acid, phenylalanine, tyrosine, glutamic acid, serine, carbamoyl aspartic acid, palmitoyl carnitine, tetradecanoyl carnitine, acetylcarnitine, and linoleylcarnitine decreased more significantly in the PYDS group.

CONCLUSIONSComparative contents of various serum metabolites changed significantly in PQDS and PYQS model groups. Some potential small molecular biomarkers related to PDS were preliminarily identified. These results might provide some data reference for exploring scientific connotation and pathological mechanisms of PDS.

Animals ; Biomarkers ; blood ; Chromatography, Liquid ; Discriminant Analysis ; Disease Models, Animal ; Drugs, Chinese Herbal ; Least-Squares Analysis ; Male ; Mass Spectrometry ; Metabolome ; Metabolomics ; Qi ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Yang Deficiency ; blood

Objective To observe the changes of abdominal oxygen saturation in very low birth weight infants (VLBWI)with feeding intolerance (FI)within 1 4 days after birth monitored by near infrared spectroscopy (NIRS).Methods VLBWI fitting entry criteria were enrolled into this study.NIRS monitoring was carried out to detect cerebral oxygen saturation (ScO2 )and abdominal oxygen saturation (SsO2 ).Data were analyzed between FI infants and feeding tolerance (FT)infants.FI was defined as follows:gastric residual of more than 50% of the previous feeding volume;emesis or abdominal distention or both;decrease,delay or discontinuation of enteral feedings. Results 93 VLBWI were enrolled.52 cases(55.91 %)presented with FI,including 29 cases(31 .1 9%)of gastric residual increasing and 23 cases(24.73%)of emesis with or without abdominal distention within 1 4 days after birth. The levels of SsO2 and SsO2 /ScO2 showed no differences in infants with FT and with FI within 24h after birth (P >0.05).The change rates of the median of SsO2 and SsO2 /ScO2 in FT infants were similar during 1 4 days (P >0.05).While both the change rates of SsO2 and SsO2 /ScO2 were markedly decreased 1 day before and the day of FI (P <0.01 ).The decreasing degree of SsO2 was similar between infants with gastric residual increasing and infants with emesis with or without abdominal distention[(1 6.2 ±5.1 )vs (1 7.4 ±3.6)%,t =0.733,P =0.476]. Conclusion Abdominal oxygen saturation measured by NIRS may be a useful method for infants adjusting the feeding plan.

Objective To investigate effects of different ventilation methods during pulmonary surfactant(PS) administration on cerebral oxygen metabolism in preterm infants with neonatal respiratory distress syndrome.Methods Newborns met the inclusion criteria were enrolled into this study,and they were randomly divided into manual group and mechanical group.During PS administration,the proximal end of the tracheal tube was connected to a bag valve mask device in the manual group or a mechanical ventilator in the mechanical group.Brain near infrared spectroscopy monitoring was carried out to detect the cerebral oxygen saturation(ScO2),and the mean arterial blood pressure (MABP) was simultaneously recorded.Results For all 49 preterm infants,PS was administered to preterm infants with severe respiratory distress syndrome treated with mechanical ventilation,including 24 cases of manual ventilation and 25 cases of mechanical ventilation.The left cerebral ScO2 and correlation coefficient of ScO2 and MABP(rScO2-MABP) showed no difference in both groups before PS administration.During administration,ScO2 dramatically increased in both groups [manual group:(85.88 ± 5.54) ％ vs.(77.31 ± 5.40) ％,t =5.521,P =0.000;mechanical group:(83.88 ± 3.18) ％ vs.(76.53 ±4.38)％,t =6.741,P =0.000],and gradually decreased after administration,the level of ScO2 didn't return to the baseline till the 2nd 5 minutes after PS administration [manual group:(79.25 ± 3.02) ％ vs.(77.31 ± 5.40) ％,t =1.560,P =0.220;mechanical group:(78.59 ± 3.45) ％ vs.(76.53 ± 4.38) ％,t =1.832,P =0.074].The same trend of ScO2 change rate was shown simultaneously in both groups.The rScO2-MABP markedly increased during administration in both groups (manual group:2.34 ±0.16 vs.1.86 ±0.21,t =9.022,P =0.000;mechanical group:2.12 ± 0.15 vs.1.87 ±0.21,t =4.810,P =0.000).The rScO2-MABt,in mechanical group rapidly decreased to baseline during the 1st5 minutes (1.84 ± 0.18 vs.1.87 ± 0.21,t =0.538,P =0.635) but went back to baseline in manual group during the 2nd 5 minutes(1.84 ±0.19 vs.1.86-0.21,t =0.350,P =0.809).Change rates of rScO2-MABP were markedly higher in manual group than those in mechanical group during the 1 st 5 minutes (1.15 ± 0.13 vs.1.00 ± 0.15,t =4.943,P =0.000).Conclusions ScO2 could be affected transiently by PS administration with different methods of ventilation.The effect on cerebral autoregulation in mechanical group is shorter than that in manual group.

BACKGROUND:Pathogenesis of Parkinson’s disease is not completely understood, and there is yet no effective therapy that can prevent the neurodegenerative process of the disease fundamentaly. OBJECTIVE:To explore the effects of pituitary adenylate cyclase-activating polypeptide (PACAP) on lactacystin-induced Parkinson’s disease dopaminergic PC12 cel apoptosis and its molecular mechanism. METHODS: Under induction by nerve growth factors, PC12 cels differentiated into dopaminergic neurons, and then were treated with different concentrations of lactacystin for different time. When the cel survival rate was about 50%,the concentration and action time oflactacystin were selected to establish cel models of Parkinson’s disease. In the study, there were control group, lactacystin group, PACAP1-27 group (intervention group 1) and PACAP1-27+PACAP6-27 co-intervention group (intervention group 2). Changes of cel morphology were observed under inverted microscope; cel viability was detected with MTT method; the expression of endoplasmic reticulum stress specific protein caspase-12 was detected by western blot. Then the action of PACAP1-27 and PACAP6-27 to the cytoxicity of lactacystin was observed. RESULTS AND CONCLUSION: With different concentrations and action time of lactacystin, the viability of PC12 cels presented a concentration- and time-dependent decline. When the lactacystin at 20μmol/L acted for 24 hours, the cel viability was declined by about 50%. Under same conditions of lactacystin concentration and action time (20 μmol/L, 24 hours), the cels in the lactacystin group appeared to have damaged changes, declined cel viability, and increased caspase-12 activity in comparison with the control group (P< 0.01). Compared with the lactacystin group, the cel damage was relieved and cel viability was increased significantly in the intervention group 1 as wel as the expression of caspase-12 was decreased (P < 0.01). Experimental findings in the intervention group 2 were similar to those in the lactacystin group. These results suggest that lactacystin, an ubiquitin proteasome inhibitor, can lead to cel damage; PACAP1-27 plays a protective role by regulating the above-mentioned signal pathway. As one PACAP1-27 receptor antagonist, PACAP6-27 can attenuate this effect of PACAP1-27.